Ca2+-binding protein, EF-hand superfamily [Signal transduction mechanisms];

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571   7 DLFWYLDYNKDGTLDIFELQEGLEDVgaiqsleeAKKIFTTGDVNKDGKLDFEEFMKYLKD-----HEKKMKLAFKSLDK 81
Cdd:COG5126    9 RRFDLLDADGDGVLERDDFEALFRRL--------WATLFSEADTDGDGRISREEFVAGMESlfeatVEPFARAAFDLLDT 80

                         90       100       110       120       130
                 ....*....|....*....|....*....|....*....|....*....|....*
gi 158258571  82 NNDGKIEASEIVQSLQTLGLtiSEQQAELILQSIDVDGTMTVDWNE----WRDYF 132
Cdd:COG5126   81 DGDGKISADEFRRLLTALGV--SEEEADELFARLDTDGDGKISFEEfvaaVRDYY 133

Mitochondrial carrier protein;

                          10        20        30        40        50        60        70        80
                  ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  175 WWRQLLAGGIAGAVSRTSTAPLDRLKIMMQVHGSKSDKMN--IFGGFRQMVKEGGIRSLWRGNGTNVIKIAPETAVKFWA 252
Cdd:pfam00153   5 FLASLLAGGIAGAIAVTVTYPLDVVKTRLQVQGGSGKSKGrgILDCFKKIYKEEGIRGLYKGLLPNLLRVAPAAAIYFGT 84

                          90
                  ....*....|..
gi 158258571  253 YEQYKKLLTEEG 264
Cdd:pfam00153  85 YETLKRLLLKKL 96

EF-hand, calcium binding motif, found in homologs of mammalian apoptosis-linked gene 2 protein (ALG-2); The family includes some homologs of mammalian apoptosis-linked gene 2 protein (ALG-2) mainly found in lower eukaryotes, such as a parasitic protist Leishmarua major and a cellular slime mold Dictyostelium discoideum. These homologs contains five EF-hand motifs. Due to the presence of unfavorable residues at the Ca2+-coordinating positions, their non-canonical EF4 and EF5 hands may not bind Ca2+. Two Dictyostelium PEF proteins are the prototypes of this family. They may bind to cytoskeletal proteins and/or signal-transducing proteins localized to detergent-resistant membranes named lipid rafts, and occur as monomers or weak homo- or heterodimers like ALG-2. They can serve as a mediator for Ca2+ signaling-related Dictyostehum programmed cell death (PCD).

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  13 DYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFM---KYLKDhekkMKLAFKSLDKNNDGKIEA 89
Cdd:cd16185   10 DRDRSGSIDVNELQKALAGGGLLFSLATAEKLIRMFDRDGNGTIDFEEFAalhQFLSN----MQNGFEQRDTSRSGRLDA 85

                         90       100       110
                 ....*....|....*....|....*....|....*....
gi 158258571  90 SEIVQSLQTLGLTISEQQAELILQSIDVDGTMTVDWNEW 128
Cdd:cd16185   86 NEVHEALAASGFQLDPPAFQALFRKFDPDRGGSLGFDDY 124

EF-hand domain pair;

                          10        20        30        40        50        60
                  ....*....|....*....|....*....|....*....|....*....|....*....|....*..
gi 158258571   69 EKKMKLAFKSLDKNNDGKIEASEIVQSLQTL--GLTISEQQAELILQSIDVDGTMTVDWNEWRDYFL 133
Cdd:pfam13499   1 EEKLKEAFKLLDSDGDGYLDVEELKKLLRKLeeGEPLSDEEVEELFKEFDLDKDGRISFEEFLELYS 67

EF-hand, calcium binding motif; EF-hands are calcium-binding motifs that occur at least in pairs. Links between disease states and genes encoding EF-hands, particularly the S100 subclass, are emerging. Each motif consists of a 12 residue loop flanked on either side by a 12 residue alpha-helix. EF-hands undergo a conformational change unpon binding calcium ions.

                           10        20
                   ....*....|....*....|....*..
gi 158258571    40 EAKKIFTTGDVNKDGKLDFEEFMKYLK 66
Cdd:smart00054   1 ELKEAFRLFDKDGDGKIDFEEFKDLLK 27

Mitochondrial carrier protein;

                          10        20        30        40        50        60        70        80
                  ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  175 WWRQLLAGGIAGAVSRTSTAPLDRLKIMMQVHGSKSDKMN--IFGGFRQMVKEGGIRSLWRGNGTNVIKIAPETAVKFWA 252
Cdd:pfam00153   5 FLASLLAGGIAGAIAVTVTYPLDVVKTRLQVQGGSGKSKGrgILDCFKKIYKEEGIRGLYKGLLPNLLRVAPAAAIYFGT 84

                          90
                  ....*....|..
gi 158258571  253 YEQYKKLLTEEG 264
Cdd:pfam00153  85 YETLKRLLLKKL 96

Mitochondrial carrier protein;

                          10        20        30        40        50        60        70        80
                  ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  364 PGVMVLLGCGALSSTCGQLASYPLALVRTRMQAQAMLEGSPQLNMVGLFRRIISKEGIPGLYRGITPNFMKVLPAVGISY 443
Cdd:pfam00153   3 LSFLASLLAGGIAGAIAVTVTYPLDVVKTRLQVQGGSGKSKGRGILDCFKKIYKEEGIRGLYKGLLPNLLRVAPAAAIYF 82

                          90
                  ....*....|.
gi 158258571  444 VVYENMKQTLG 454
Cdd:pfam00153  83 GTYETLKRLLL 93

EF-hand, calcium binding motif, found in homologs of mammalian apoptosis-linked gene 2 protein (ALG-2); The family includes some homologs of mammalian apoptosis-linked gene 2 protein (ALG-2) mainly found in lower eukaryotes, such as a parasitic protist Leishmarua major and a cellular slime mold Dictyostelium discoideum. These homologs contains five EF-hand motifs. Due to the presence of unfavorable residues at the Ca2+-coordinating positions, their non-canonical EF4 and EF5 hands may not bind Ca2+. Two Dictyostelium PEF proteins are the prototypes of this family. They may bind to cytoskeletal proteins and/or signal-transducing proteins localized to detergent-resistant membranes named lipid rafts, and occur as monomers or weak homo- or heterodimers like ALG-2. They can serve as a mediator for Ca2+ signaling-related Dictyostehum programmed cell death (PCD).

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  13 DYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFM---KYLKDhekkMKLAFKSLDKNNDGKIEA 89
Cdd:cd16185   10 DRDRSGSIDVNELQKALAGGGLLFSLATAEKLIRMFDRDGNGTIDFEEFAalhQFLSN----MQNGFEQRDTSRSGRLDA 85

                         90       100       110
                 ....*....|....*....|....*....|....*....
gi 158258571  90 SEIVQSLQTLGLTISEQQAELILQSIDVDGTMTVDWNEW 128
Cdd:cd16185   86 NEVHEALAASGFQLDPPAFQALFRKFDPDRGGSLGFDDY 124

ADP/ATP transporter on adenylate translocase; Provisional

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571 160 TIPDEFTEDEKKSgQWWR----QLLAGGIAGAVSRTSTAPLD--RLKIMMQVhGSKSDK--MNIFGGFRQMVKEGGIRSL 231
Cdd:PTZ00169  95 YFKNMFPKYNQKT-DFWKffgvNILSGGLAGASSLLIVYPLDfaRTRLASDI-GKGGDRefTGLFDCLMKISKQTGFLSL 172

                         90       100       110       120       130       140       150       160
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571 232 WRGNGTNVIKIAPETAVKFWAYEQYKKLLTEEGQKIGTFERFISGSMAGATAQTFIYPMEVMKTRLAV------GKTGQY 305
Cdd:PTZ00169 173 YQGFGVSVQGIIVYRGAYFGLYDSAKALLFGNDKNTNILYKWAVAQTVTILAGLISYPFDTVRRRMMMmsgrkaKSEIQY 252

                        170       180
                 ....*....|....*....|....*...
gi 158258571 306 SGIYDCAKKILKHEGLGAFYKGYVPNLL 333
Cdd:PTZ00169 253 TGTLDCWKKILKNEGLGGFFKGAWANVL 280

calmodulin; Provisional

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571   5 YGDLFWYLDYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFMKYLK------DHEKKMKLAFKS 78
Cdd:PTZ00184  13 FKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTLMArkmkdtDSEEEIKEAFKV 92

                         90       100       110       120       130
                 ....*....|....*....|....*....|....*....|....*....|
gi 158258571  79 LDKNNDGKIEASEIVQSLQTLGLTISEQQAELILQSIDVDGTMTVDWNEW 128
Cdd:PTZ00184  93 FDRDGNGFISAAELRHVMTNLGEKLTDEEVDEMIREADVDGDGQINYEEF 142

EF-hand, calcium binding motif; A diverse superfamily of calcium sensors and calcium signal modulators; most examples in this alignment model have 2 active canonical EF hands. Ca2+ binding induces a conformational change in the EF-hand motif, leading to the activation or inactivation of target proteins. EF-hands tend to occur in pairs or higher copy numbers.

                         10        20        30        40        50
                 ....*....|....*....|....*....|....*....|....*....|....*
gi 158258571  12 LDYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFMKYLK 66
Cdd:cd00051    9 FDKDGDGTISADELKAALKSLGEGLSEEEIDEMIREVDKDGDGKIDFEEFLELMA 63

EF-hand domain pair;

                          10        20        30        40        50        60
                  ....*....|....*....|....*....|....*....|....*....|....*....|....*..
gi 158258571   69 EKKMKLAFKSLDKNNDGKIEASEIVQSLQTL--GLTISEQQAELILQSIDVDGTMTVDWNEWRDYFL 133
Cdd:pfam13499   1 EEKLKEAFKLLDSDGDGYLDVEELKKLLRKLeeGEPLSDEEVEELFKEFDLDKDGRISFEEFLELYS 67

EF-hand domain pair;

                          10        20        30        40        50        60
                  ....*....|....*....|....*....|....*....|....*....|....*....|..
gi 158258571    7 DLFWYLDYNKDGTLDIFELQEGLEDVGAIQSL--EEAKKIFTTGDVNKDGKLDFEEFMKYLK 66
Cdd:pfam13499   6 EAFKLLDSDGDGYLDVEELKKLLRKLEEGEPLsdEEVEELFKEFDLDKDGRISFEEFLELYS 67

EF-hand, calcium binding motif, found in uncharacterized calglandulin-like proteins; The family corresponds to a group of uncharacterized calglandulin-like proteins. Although their biological function remain unclear, they show high sequence similarity with human calglandulin-like protein GAGLP, which is an ortholog of calglandulin from the venom glands of Bothrops insularis snake. Both GAGLP and calglandulin are putative Ca2+-binding proteins with four EF-hand motifs. However, members in this family contain only three EF-hand motifs. In this point, they may belong to the parvalbumin-like EF-hand family, which is characterized by the presence of three consecutive EF-hand motifs (helix-loop-helix).

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  49 DVNKDGKLDFEEFMKYLK------DHEKKMKLAFKSLDKNNDGKIEASEIVQSLQTLGLT-----ISEQQAELILQSIDV 117
Cdd:cd16252   10 EMRHHGSFNYSKFFEYMQkfqtseQQEEAIRKAFQMLDKDKSGFIEWNEIKYILSTVPSSmpvapLSDEEAEAMIQAADT 89

                         90
                 ....*....|...
gi 158258571 118 DGTMTVDWNEWRD 130
Cdd:cd16252   90 DGDGRIDFQEFSD 102

EF-hand, calcium binding motif, found in reticulocalbin-2 (RCN2) mainly from protostomes; This family corresponds to a group of uncharacterized RCN2-like proteins, which are mainly found in protostomes. Although their biological function remains unclear, they show high sequence similarity with RCN2 (also known as E6BP or TCBP-49), which is an endoplasmic reticulum resident low-affinity Ca2+-binding protein that has been implicated in immunity, redox homeostasis, cell cycle regulation and coagulation. Members in this family contain six copies of the EF-hand Ca2+-binding motif, but may lack a C-terminal His-Asp-Glu-Leu (HDEL) tetrapeptide that is required for retention of RCN2 in the endoplasmic reticulum (ER).

                         10        20        30        40        50        60
                 ....*....|....*....|....*....|....*....|....*....|....*....|....
gi 158258571  36 QSLEEAKKIFTTGDVNKDGKLDFEEFMKYLKDHE-KKM-----KLAFKSLDKNNDGKIEASEIV 93
Cdd:cd16227  119 KLLEDDKEMFEAADLNKDGKLDKTEFSAFQHPEEyPHMhpvliEQTLRDKDKDNDGFISFQEFL 182

Penta-EF hand, calcium binding motifs, found in calcium-dependent protease small subunit CAPNS1 and CAPNS2; CAPNS1, also termed calpain small subunit 1 (CSS1), or calcium-activated neutral proteinase small subunit (CANP small subunit), or calcium-dependent protease small subunit (CDPS), or calpain regulatory subunit, is a common 28-kDa regulatory calpain subunit encoded by the calpain small 1 (Capns1, also known as Capn4) gene. It acts as a binding partner to form a heterodimer with the 80 kDa calpain large catalytic subunit and is required in maintaining the activity of calpain. CAPNS1 plays a significant role in tumor progression of human cancer, and functions as a potential therapeutic target in human hepatocellular carcinoma (HCC), nasopharyngeal carcinoma (NPC), glioma, and clear cell renal cell carcinoma (ccRCC). It may be involved in regulating migration and cell survival through binding to the SH3 domain of Ras GTPase-activating protein (RasGAP). It may also modulate Akt/FoxO3A signaling and apoptosis through PP2A. CAPNS1 contains an N-terminal glycine rich domain and a C-terminal PEF-hand domain. CAPNS2, also termed calpain small subunit 2 (CSS2), is a novel tissue-specific 30 kDa calpain small subunit that lacks two oligo-Gly stretches characteristic of the N-terminal Gly-rich domain of CAPNS1. CAPNS2 acts as a chaperone for the calpain large subunit, and appears to be the functional equivalent of CAPNS1. However, CAPNS2 binds the large subunit much more weakly than CAPNS1 and it does not undergo the autolytic conversion typical of CAPNS1.

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  37 SLEEAKKIFTTGDVNKDGKLDFEEFmKYLKDHEKKMKLAFKSLDKNNDGKIEASEIVQSLQTLGLTISEQQAELILQSI- 115
Cdd:cd16188   41 GIDTCRSMVAVMDSDTTGKLGFEEF-KYLWNNIKKWQGIYKQFDTDRSGTIGSQELPGAFEAAGFHLNEQLYQMIIRRYs 119

                 ....*....
gi 158258571 116 DVDGTMTVD 124
Cdd:cd16188  120 DEDGNMDFD 128

EF-hand domain pair;

                          10        20        30        40        50        60
                  ....*....|....*....|....*....|....*....|....*....|....*....|...
gi 158258571   39 EEAKKIFTTGDVNKDGKLDFEEFMKYLKDHEKKMKLA-------FKSLDKNNDGKIEASEIVQ 94
Cdd:pfam13499   2 EKLKEAFKLLDSDGDGYLDVEELKKLLRKLEEGEPLSdeeveelFKEFDLDKDGRISFEEFLE 64

EF-hand, calcium binding motif, found in 45 kDa calcium-binding protein (Cab45); Cab45, also termed stromal cell-derived factor 4 (SDF-4), is a soluble, lumenal Golgi resident low-affinity Ca2+-binding protein that contains six copies of the EF-hand Ca2+-binding motif. It is required for secretory pathway calcium ATPase1 (SPCA1)-dependent Ca2+ import into the trans-Golgi network (TGN) and plays an essential role in Ca2+-dependent secretory cargo sorting at the TGN.

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  48 GDVNKDGKLDFEEFMKylkdheKKMKLAFKSLDKNNDGKIEASE----IVQSLQTlGLTISEQQAELILQSIDVDGTMTV 123
Cdd:cd16225   18 GNEKEEFEEDSEPKKR------KKLKEIFKKVDVNTDGFLSAEEledwIMEKTQE-HFQEAVEENEQIFKAVDTDKDGNV 90

                         90
                 ....*....|
gi 158258571 124 DWNEWRDYFL 133
Cdd:cd16225   91 SWEEYRVHFL 100

Penta-EF hand, calcium binding motifs, found in calpain-9 (CAPN9); CAPN9, also termed digestive tract-specific calpain, or new calpain 4 (nCL-4), or protein CG36, is a calpain large subunit predominantly expressed in gastrointestinal tract. It plays a physiological role in the suppression of tumorigenesis. It acts as an important biomolecule link for the regression of colorectal cancer via intracellular calcium homeostasis. CAPN9 may also play a critical role in lumen formation. Moreover, CAPN9, together with CAPN8, forms an active protease complex, G-calpain, in which both proteins are essential for stability and activity. The G-Calpain has been implicated in gastric mucosal defense. Furthermore, down-regulation of calpain 9 has been linked to hypertensive heart and kidney disease in salt-sensitive Dahl rats. CAPN9 contains a short N-terminal anchor helix, followed by a calpain cysteine protease (CysPc) domain, a C2-domain-like (C2L) domain, and a C-terminal Ca2+-binding penta-EF-hand (PEF) domain.

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  16 KDGTLDIFELQEGLEDVGAIQ--------SLEEAKKIFTTGDVNKDGKLDFEEFmKYLKDHEKKMKLAFKSLDKNNDGKI 87
Cdd:cd16192   12 EDLEISAEELQYVLNAVLARTkeikfkklSLLSCKNIISLMDTSGNGKLGFSEF-KVFWDKLKKWIGLFLKYDADRSGTM 90

                         90       100
                 ....*....|....*....|....
gi 158258571  88 EASEIVQSLQTLGLTISEQQAELI 111
Cdd:cd16192   91 SSYELRSALKAAGFQLNNQLLQLI 114

XopAW family type III secretion system calcium-binding effector;

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571   7 DLFWYLDYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFMKYLK------DHEKKMKLA---FK 77
Cdd:NF041410  31 QLFAKLDSDGDGSVSQDELSSALSSKSDDGSLIDLSELFSDLDSDGDGSLSSDELAAAAPppppppDQAPSTELAddlLS 110

                         90       100       110       120
                 ....*....|....*....|....*....|....*....|....*.
gi 158258571  78 SLDKNNDGKIEASEIVQSLQTLGltiSEQQAELILQSIDVDGTMTV 123
Cdd:NF041410 111 ALDTDGDGSISSDELSAGLTSAG---SSADSSQLFSALDSDGDGSV 153

EF-hand motif found in eukaryotic phosphoinositide-specific phospholipase C (PI-PLC, EC 3.1.4.11) isozymes; PI-PLC isozymes are signaling enzymes that hydrolyze the membrane phospholipids phosphatidylinositol-4,5-bisphosphate (PIP2) to generate two important second messengers in eukaryotic signal transduction cascades, Inositol 1,4,5-trisphosphate (InsP3) and diacylglycerol (DAG). InsP3 triggers inflow of calcium from intracellular stores, while DAG, together with calcium, activates protein kinase C, which goes on to phosphorylate other molecules, leading to altered cellular activity. Calcium is required for the catalysis. This family corresponds to the four EF-hand motifs containing PI-PLC isozymes, including PI-PLC-beta (1-4), -gamma (1-2), -delta (1,3,4), -epsilon (1), -zeta (1), eta (1-2). Lower eukaryotes such as yeast and slime molds contain only delta-type isozymes. In contrast, other types of isoforms present in higher eukaryotes. This family also includes 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase 1 (PLC1) from fungi. Some homologs from plants contain only two atypical EF-hand motifs and they are not included. All PI-PLC isozymes except sperm-specific PI-PLC-zeta share a core set of domains, including an N-terminal pleckstrin homology (PH) domain, four atypical EF-hand motifs, a PLC catalytic core, and a single C2 domain. PI-PLC-zeta lacks the PH domain. The PLC catalytic core domain is a TIM barrel with two highly conserved regions (X and Y) split by a highly degenerate linker sequence. Most of EF-hand motifs found in PI-PLCs consist of a helix-loop-helix structure, but lack residues critical to metal binding. Moreover, the EF-hand region of most of PI-PLCs may have an important regulatory function, but it has yet to be identified. However, PI-PLC-zeta is a key exception. It is responsible for Ca2+ oscillations in fertilized oocytes and exhibits a high sensitivity to Ca2+ mediated through its EF-hand domain. In addition, PI-PLC-eta2 shows a canonical EF-loop directing Ca2+-sensitivity and thus can amplify transient Ca2+ signals. Also it appears that PI-PLC-delta1 can regulate the binding of PH domain to PIP2 in a Ca2+-dependent manner through its functionally important EF-hand domains. PI-PLCs can be activated by a variety of extracellular ligands, such as growth factors, hormones, cytokines and lipids. Their activation has been implicated in tumorigenesis and/or metastasis linked to migration, proliferation, growth, inflammation, angiogenesis and actin cytoskeleton reorganization. PI-PLC-beta isozymes are activated by G-protein coupled receptor (GPCR) through different mechanisms. However, PI-PLC-gamma isozymes are activated by receptor tyrosine kinase (RTK), such as Rho and Ras GTPases. In contrast, PI-PLC-epsilon are activated by both GPCR and RTK. PI-PLC-delta1 and PLC-eta 1 are activated by GPCR-mediated calcium mobilization. The activation mechanism for PI-PLC-zeta remains unclear.

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571   9 FWYL-DYNKDGTLDIFELQEGLEDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEF---MKYLKDHeKKMKLAFKSLDKNND 84
Cdd:cd15898    5 QWIKaDKDGDGKLSLKEIKKLLKRLNIRVSEKELKKLFKEVDTNGDGTLTFDEFeelYKSLTER-PELEPIFKKYAGTNR 83

                         90       100       110
                 ....*....|....*....|....*....|....*..
gi 158258571  85 GKIEASEIVQSLQ-TLGLTISEQQAELILQSIDVDGT 120
Cdd:cd15898   84 DYMTLEEFIRFLReEQGENVSEEECEELIEKYEPERE 120

EF-hand, calcium binding motif, found in the hexa-EF hand proteins family; The hexa-EF hand proteins family, also named the calbindin sub-family, contains a group of six EF-hand Ca2+-binding proteins, including calretinin (CR, also termed 29 kDa calbindin), calbindin D28K (CB, also termed vitamin D-dependent calcium-binding protein, avian-type), and secretagogin (SCGN). CR is a cytosolic hexa-EF-hand calcium-binding protein predominantly expressed in a variety of normal and tumorigenic t-specific neurons of the central and peripheral nervous system. It is a multifunctional protein implicated in many biological processes, including cell proliferation, differentiation, and cell death. CB is highly expressed in brain tissue. It is a strong calcium-binding and buffering protein responsible for preventing a neuronal death as well as maintaining and controlling calcium homeostasis. SCGN is a six EF-hand calcium-binding protein expressed in neuroendocrine, pancreatic endocrine and retinal cells. It plays a crucial role in cell apoptosis, receptor signaling and differentiation. It is also involved in vesicle secretion through binding to various proteins, including interacts with SNAP25, SNAP23, DOC2alpha, ARFGAP2, rootletin, KIF5B, beta-tubulin, DDAH-2, ATP-synthase and myeloid leukemia factor 2. SCGN functions as a Ca2+ sensor/coincidence detector modulating vesicular exocytosis of neurotransmitters, neuropeptides or hormones. Although the family members share a significant amount of secondary sequence homology, they display altered structural and biochemical characteristics, and operate in distinct fashions. CB contains six EF-hand motifs in a single globular domain, where EF-hands 1, 3, 4, 5 bind four calcium ions. CR contains six EF-hand motifs within two independent domains, CR I-II and CR III-VI. They harbor two and four EF-hand motifs, respectively. The first 5 EF-hand motifs are capable of binding calcium ions, while the EF-hand 6 is inactive. SCGN consists of the three globular domains each of which contains a pair of EF-hand motifs. Human SCGN simultaneously binds four calcium ions through its EF-hands 3, 4, 5 and 6 in one high affinity and three low affinity calcium-binding sites. In contrast, SCGNs in other lower eukaryotes, such as D. rerio, X. laevis, M. domestica, G. gallus, O. anatinus, are fully competent in terms of six calcium-binding.

                         10        20        30        40        50        60        70        80
                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  12 LDYNKDGTLDIFELQEGL----------EDVGAIQSLEEAKKIFTTGDVNKDGKLDFEEFMKYLKDHEKKMK-------- 73
Cdd:cd15902   53 YDENEDGKIEIRELANILpteenflllfRREQPLISSVEFMKIWRKYDTDGSGFIEAKELKGFLKDLLLKNKkhvsppkl 132

                         90       100
                 ....*....|....*....|....*...
gi 158258571  74 -----LAFKSLDKNNDGKIEASEIVQSL 96
Cdd:cd15902  133 deytkLILKEFDANKDGKLELDEMAKLL 160

Penta-EF hand, calcium binding motifs, found in Drosophila melanogaster calpain-A (CalpA), calpain-B (CalpB), and similar proteins; The family contains two calpains that have been found in Drosophila, CalpA and CalpB. CalpA, also termed calcium-activated neutral proteinase A (CANP A), or calpain-A catalytic subunit, is a Drosophila calpain homolog specifically expressed in a few neurons in the central nervous system, in scattered endocrine cells in the midgut, and in blood cells. CalpB, also termed calcium-activated neutral proteinase B (CANP B), contains calpain-B catalytic subunit 1 and calpain-B catalytic subunit 2. Both CalpA and CalpB are closely related to that of vertebrate calpains, and they share similar domain architecture, which consists of four domains: the N-terminal domain I, the catalytic domain II carrying the three active site residues, Cys, His and Asn, the Ca2+-regulated phospholipid-binding domain III, and penta-EF-hand Ca2+-binding domain IV. Besides, CalpA and CalpB display some distinguishing structural features that are not found in mammalian typical calpains. CalpA harbors a 76 amino acid long hydrophobic stretch inserted in domain IV, which may be involved in membrane attachment of this enzyme. CalpB has an unusually long N-terminal tail of 224 amino acids, which belongs to the class of intrinsically unstructured proteins (IUP) and may become ordered upon binding to target protein(s). Moreover, they do not need small regulatory subunits for their catalytic activity, and their proteolytic function is not regulated by an intrinsic inhibitor as the Drosophila genome contains neither regulatory subunit nor calpastatin orthologs. As a result, they may exist as a monomer or perhaps as a homo- or heterodimer together with a second large subunit. Furthermore, both CalpA and CalpB are dispensable for viability and fertility and do not share vital functions during Drosophila development. Phosphatidylinositol 4,5-diphosphate, phosphatidylinositol 4-monophosphate, phosphatidylinositol, and phosphatidic acid can stimulate the activity and the rate of activation of CalpA, but not CalpB. Calpain A modulates Toll responses by limited Cactus/IkappaB proteolysis. CalpB directly interacts with talin, an important component of the focal adhesion complex, and functions as an important modulator in border cell migration within egg chambers, which may act via the digestion of talin. CalpB can be phosphorylated by cAMP-dependent protein kinase (protein kinase A, PKA; EC 2.7.11.11) at Ser240 and Ser845, as well as by mitogen-activated protein kinase (ERK1 and ERK2; EC 2.7.11.24) at Thr747. The activation of the ERK pathway by extracellular signals results in the phosphorylation and activation of calpain B. In Schneider cells (S2), calpain B was mainly in the cytoplasm and upon a rise in Ca2+ the enzyme adhered to intracellular membranes.

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  16 KDGTLDIFELQEGLEDV------GAIQSLEEAKKIFTTGDVNKDGKLDFEEFMKyLKDHEKKMKLAFKSLDKNNDGKIEA 89
Cdd:cd16196   12 EDMEIDAYELQDILNTAfkkdfpFDGFSLDACRSMVAMMDVDRSGKLGFEEFKK-LWEDLRSWKRVFKLFDTDGSGSFSS 90

                         90
                 ....*....|....*
gi 158258571  90 SEIVQSLQTLGLTIS 104
Cdd:cd16196   91 FELRNALNSAGFRLS 105

EF-hand motif found in eukaryotic phosphoinositide-specific phospholipase C (PI-PLC, EC 3.1.4.11) isozymes; PI-PLC isozymes are signaling enzymes that hydrolyze the membrane phospholipids phosphatidylinositol-4,5-bisphosphate (PIP2) to generate two important second messengers in eukaryotic signal transduction cascades, Inositol 1,4,5-trisphosphate (InsP3) and diacylglycerol (DAG). InsP3 triggers inflow of calcium from intracellular stores, while DAG, together with calcium, activates protein kinase C, which goes on to phosphorylate other molecules, leading to altered cellular activity. Calcium is required for the catalysis. This family corresponds to the four EF-hand motifs containing PI-PLC isozymes, including PI-PLC-beta (1-4), -gamma (1-2), -delta (1,3,4), -epsilon (1), -zeta (1), eta (1-2). Lower eukaryotes such as yeast and slime molds contain only delta-type isozymes. In contrast, other types of isoforms present in higher eukaryotes. This family also includes 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase 1 (PLC1) from fungi. Some homologs from plants contain only two atypical EF-hand motifs and they are not included. All PI-PLC isozymes except sperm-specific PI-PLC-zeta share a core set of domains, including an N-terminal pleckstrin homology (PH) domain, four atypical EF-hand motifs, a PLC catalytic core, and a single C2 domain. PI-PLC-zeta lacks the PH domain. The PLC catalytic core domain is a TIM barrel with two highly conserved regions (X and Y) split by a highly degenerate linker sequence. Most of EF-hand motifs found in PI-PLCs consist of a helix-loop-helix structure, but lack residues critical to metal binding. Moreover, the EF-hand region of most of PI-PLCs may have an important regulatory function, but it has yet to be identified. However, PI-PLC-zeta is a key exception. It is responsible for Ca2+ oscillations in fertilized oocytes and exhibits a high sensitivity to Ca2+ mediated through its EF-hand domain. In addition, PI-PLC-eta2 shows a canonical EF-loop directing Ca2+-sensitivity and thus can amplify transient Ca2+ signals. Also it appears that PI-PLC-delta1 can regulate the binding of PH domain to PIP2 in a Ca2+-dependent manner through its functionally important EF-hand domains. PI-PLCs can be activated by a variety of extracellular ligands, such as growth factors, hormones, cytokines and lipids. Their activation has been implicated in tumorigenesis and/or metastasis linked to migration, proliferation, growth, inflammation, angiogenesis and actin cytoskeleton reorganization. PI-PLC-beta isozymes are activated by G-protein coupled receptor (GPCR) through different mechanisms. However, PI-PLC-gamma isozymes are activated by receptor tyrosine kinase (RTK), such as Rho and Ras GTPases. In contrast, PI-PLC-epsilon are activated by both GPCR and RTK. PI-PLC-delta1 and PLC-eta 1 are activated by GPCR-mediated calcium mobilization. The activation mechanism for PI-PLC-zeta remains unclear.

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  71 KMKLAFKSLDKNNDGKIEASEIVQSLQTLGLTISEQQAELILQSIDVDGTMTVDWNEWRD-----------YFLFN---- 135
Cdd:cd15898    1 WLRRQWIKADKDGDGKLSLKEIKKLLKRLNIRVSEKELKKLFKEVDTNGDGTLTFDEFEElykslterpelEPIFKkyag 80

                         90
                 ....*....|....*..
gi 158258571 136 ---PVTDIEEIIRFWKH 149
Cdd:cd15898   81 tnrDYMTLEEFIRFLRE 97

EF-hand motif found in phosphoinositide phospholipase C delta 1 (PI-PLC-delta1); PI-PLC-delta1, also termed 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase delta-1 (PLCD1), or phospholipase C-III (PLC-III), or phospholipase C-delta-1 (PLC-delta-1), is present in high abundancy in the brain, heart, lung, skeletal muscle and testis. It is activated by high calcium levels generated by other PI-PLC family members, and therefore functions as a calcium amplifier within the cell. PI-PLC-delta1 is required for maintenance of homeostasis in skin and metabolic tissues. Moreover, it is essential in trophoblasts for placental development. Simultaneous loss of PI-PLC-delta1 may cause placental vascular defects, leading to embryonic lethality. PI-PLC-delta1 can be positively or negatively regulated by several binding partners, including p122/Rho GTPase activating protein (RhoGAP), Gha/Transglutaminase II, RalA, and calmodulin. It is involved in Alzheimer's disease and hypertension. Furthermore, PI-PLC-delta1 regulates cell proliferation and cell-cycle progression from G1- to S-phase by control of cyclin E-CDK2 activity and p27 levels. It can be activated by alpha1-adrenoreceptors (AR) in a calcium-dependent manner and may be important for G protein-coupled receptors (GPCR) responses in vascular smooth muscle (VSM). PI-PLC-delta1 may also be involved in noradrenaline (NA)-induced phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis and modulate sustained contraction of mesenteric small arteries. In addition, it inhibits thermogenesis and induces lipid accumulation, and therefore contributes to the development of obesity. PI-PLC-delta1 contains a core set of domains, including an N-terminal pleckstrin homology (PH) domain, four atypical EF-hand motifs, a PLC catalytic core, and a single C-terminal C2 domain. The PLC catalytic core domain is a TIM barrel with two highly conserved regions (X and Y) split by a highly degenerate linker sequence. PI-PLC-delta1 can regulate the binding of PH domain to PIP2 in a Ca2+-dependent manner through its functionally important EF-hand domains. In addition, PI-PLC-delta1 possesses a classical leucine-rich nuclear export sequence (NES) located in the EF hand motifs, as well as a nuclear localization signal within its linker region, both of which may be responsible for translocating PI-PLC-delta1 into and out of the cell nucleus.

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  49 DVNKDGKLDFEEFMKYLKD-----HEKKMKLAFKSLDKNNDGKIEASEIvQSLQTLgLTISEQQAELILQSIDVDGTMTV 123
Cdd:cd16217   10 DKNKDNKMSFKELKDFLKEinievDDDYAEKLFKECDKSKSGFLEGEEI-EEFYKL-LTKREEIDVIFGEYAKSDGTMSR 87

                 .
gi 158258571 124 D 124
Cdd:cd16217   88 N 88

EF-hand, calcium binding motif, found in 45 kDa calcium-binding protein (Cab45); Cab45, also termed stromal cell-derived factor 4 (SDF-4), is a soluble, lumenal Golgi resident low-affinity Ca2+-binding protein that contains six copies of the EF-hand Ca2+-binding motif. It is required for secretory pathway calcium ATPase1 (SPCA1)-dependent Ca2+ import into the trans-Golgi network (TGN) and plays an essential role in Ca2+-dependent secretory cargo sorting at the TGN.

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571   7 DLFWYLDYNKDGTLDIFEL--------QEGLEDvgaiqSLEEAKKIFTTGDVNKDGKLDFEEF-MKYLKDHEKKMKLAFK 77
Cdd:cd16225   38 EIFKKVDVNTDGFLSAEELedwimektQEHFQE-----AVEENEQIFKAVDTDKDGNVSWEEYrVHFLLSKGYSEEEAEE 112

                         90
                 ....*....|....*
gi 158258571  78 SLDKNNDGKIEASEI 92
Cdd:cd16225  113 KIKNNEELKLDEDDK 127

EF-hand, calcium binding motif, found in reticulocalbin-2 (RCN2) mainly from protostomes; This family corresponds to a group of uncharacterized RCN2-like proteins, which are mainly found in protostomes. Although their biological function remains unclear, they show high sequence similarity with RCN2 (also known as E6BP or TCBP-49), which is an endoplasmic reticulum resident low-affinity Ca2+-binding protein that has been implicated in immunity, redox homeostasis, cell cycle regulation and coagulation. Members in this family contain six copies of the EF-hand Ca2+-binding motif, but may lack a C-terminal His-Asp-Glu-Leu (HDEL) tetrapeptide that is required for retention of RCN2 in the endoplasmic reticulum (ER).

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*
gi 158258571  59 EEFmKYLKDHEKKMKLA--FKSLDKNNDGKIEASE----IVQSLQTLgltiSEQQAELILQSIDVDGTMTVDWNE 127
Cdd:cd16227   24 EEF-DELPPEEAKRRLAvlAKKMDLNDDGFIDRKElkawILRSFKML----DEEEANERFEEADEDGDGKVTWEE 93

phosphatidylserine decarboxylase

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                 ....*....|....*....|....*....|....*....|....*....|....*....|....*....|....*....|
gi 158258571  41 AKKIFTTGDVNKDGKLDFEEFMKYLKDH------EKKMKLaFKSLDKNNDGKIEASEIVQSLQTlgltisEQQAELILQS 114
Cdd:PLN02964 181 ARRILAIVDYDEDGQLSFSEFSDLIKAFgnlvaaNKKEEL-FKAADLNGDGVVTIDELAALLAL------QQEQEPIINN 253

                 ....*
gi 158258571 115 IDVDG 119
Cdd:PLN02964 254 CPVCG 258