Skip to main content
Log in

Classification and evolution of EF-hand proteins

  • Published:
Biometals Aims and scope Submit manuscript

Abstract

Forty-five distinct subfamilies of EF-hand proteins have been identified. They contain from two to eight EF-hands that are recognizable by amino acid sequence as being statistically similar to other EF-hand domains. All proteins within one subfamily are congruent to one another, i.e. the dendrogram computed from one of the EF-hand domains is similar, within statistical error, to the dendrogram computed from another(s) domain. Thirteen subfamilies - including Calmodulin, Troponin C, Essential light chain, Regulatory light chain - referred to collectively as CTER, are congruent with one another. They appear to have evolved from a single ur-domain by two cycles of gene duplication and fusion. The subfamilies of CTER subsequently evolved by gene duplications and speciations. The remaining 32 subfamilies do not show such general patterns of congruence; however, some - such as S100, intestinal calcium binding protein (calbindin 9kd), and trichohylin - do not form congruent clusters of subfamilies. Nearly all of the domains 1, 3, 5, and 7 are most similar to other ODD domains. Correspondingly the EVEN numbered domains of all 45 subfamilies most closely resemble EVEN domains of other subfamilies. Many sequence and chem-ical characteristics do not show systemic trends by subfamily or species of host organisms; such homoplasy is widespread. Eighteen of the subfamilies are heterochimeric; in addition to multiple EF-hands they contain domains of other evolutionary origins.© Kluwer Academic Publishers

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Adachi J, Hasegawa M. 1996 MOLPHY Version 2.3: Programs for molecular phylogenetics based on maximum likelihood. Computer Science Monographs, 28, 1–150. Institute of Statistical Mathematics, Tokyo.

    Google Scholar 

  • Ajioka J, Swindle J. 1993 The calmodulin-ubiquitin associated genes of Trypanosoma cruzi: their identification and transcription. Mol. Biochem. Parasitol. 57, 127–136.

    Google Scholar 

  • Altschul SF, Boguski MS, Gish W, Wootton JC. 1994 Issues in searching molecular sequence databases Nature Genetics 6, 119 – 129.

    Google Scholar 

  • Ames JB, Porumb T, Tanaka T, Ikura M, Stryer L. 1995 Amino-terminal myristoylation induces cooperative calcium binding to recoverin. J. Biol. Chem. 270, 4526–4533.

    Google Scholar 

  • Barroso MB, Bernd KK, DeWitt ND, Chang A, Mills K, Stzul ES. 1996 A novel Ca2+-binding protein, p22, is required for constitutive membrane traffic. J. Biol. Chem. 271, 10183–10187.

    Google Scholar 

  • Bartling D, Bülter H. Weiler EW. 1993 Arabidopsis thaliana cDNA encoding a novel member of the EF-Hand superfamily of calcium-binding proteins. Plant Physiol. 102, 1059–1060.

    Google Scholar 

  • Bearer EL, DeGiorgis JA, Jaffe H, Medeiros NA, Reese TS. 1996 An axoplasmic myosin with a calmodulin-like light chain. Proc. Natl. Acad. Sci. USA 93, 6064–6068

    Google Scholar 

  • Berchtold MW. 1993 Evolution of EF-hand calcium-modulated proteins. V. The genes encoding EF-hand proteins are not clustered in mammalian genomes. J. Mol. Evol. 36, 489–496.

    Google Scholar 

  • Biggins S, Ros, MD. 1994 Direct interaction between yeast spindle pole body components: Kar1p is required for Cdc31p localization to the spindle pole body. J. Cell Biol. 125, 843–852.

    Google Scholar 

  • Blanchard H, Grochulski P, Li Y, Arthr SC, Davies PL, Elce JS, Cygle M. 1997 Structure of a calpain Ca2+-binding domain reveals a novel EF-hand and Ca2+-induced conformational changes. Nature Struct. Biol. 4, 532–538.

    Google Scholar 

  • Brown LJ, MacDonald MJ, Lehn DA, Moran SM. 1994 Sequence of rat mitochondrial glycerol-3-phosphate dehydrogenase cDNA. Evidence of EF-hand calcium-binding domains. J. Biol. Chem. 269, 14363–14366.

    Google Scholar 

  • Brownawell AM, Creutz CE. 1997 Calcium-dependent binding of sorcin to the N-terminal domain of synexin (annexin VII). J. Biol. Chem. 272, 22182–22190.

    Google Scholar 

  • Chazin WJ. 1998 Structural aspects of calcium-binding proteins of the EF-hand type. BioMetals this issue.

  • Chothia C. 1992 One thousand families for the molecular biologist. Nature 357, 543–544.

    Google Scholar 

  • Cook WJ, Ealik SE, Babu YS, Cox JA, Vijay Kumar S. 1991 Three-dimensional structure of a sarcoplasmic calcium-binding protein from Nereis divesicolor. J. Biol. Chem. 266, 652–656.

    Google Scholar 

  • Cook WJ, Jeffrey LC, Cox JA Vijay Kumar S. 1993 Structure of a sarcoplasmic calcium-binding protein from amphioxus refined at 2.4 Å resolution. J. Mol. Biol. 229, 461–471.

    Google Scholar 

  • Cox JA, Alard P, Schaad O. 1990 Comparative molecular modeling of Amphioxus calcium vector protein with calmodulin and troponin C. Prot. Engineering 4, 23–32.

    Google Scholar 

  • Crivici A, Ikura M. 1995 Molecular and structural basis of target recognition by calmodulin. Annu. Rev. Biophys. Biomol. Struct. 24, 85–116.

    Google Scholar 

  • de Arruda MV, Watson S, Lin C-S, Leavitt J, Matsudaira P. 1990 Fimbrin is a homologue of the cytoplasmic phosphoprotein plastin and has domains homologous with calmodulin and actin gelatin proteins. J. Cell Biol. 111, 1069–1079.

    Google Scholar 

  • Deka N, Wong E, Matera AG, Kraft R, Leinwand LA, Schmid CW. 1988 Repetitive nucleotide sequence insertions into a novel calmodulin-related gene and its processed pseudogene. Gene 71, 123–134.

    Google Scholar 

  • Donaldson C, Barber KR, Kay CM, Shaw GS. 1995 Human S100b protein: Formation of a tetramer from synthetic calcium-binding site peptides. Protein Sci. 4, 765–772.

    Google Scholar 

  • Engman DM, Krause K-H, Blumin JH, Kim KS, Kirchhoff LV, Donelson, JE. 1989 A novel flagellar Ca2+-binding protein of trypanosomes. J. Biol. Chem. 264, 18627–18631.

    Google Scholar 

  • Essen L-O, Perisic O, Cheung R, Katan M, Williams RL. 1996 Crystal structure of a mammalian phosphoinositide specific phospholipase Cδ. Nature 380, 595–602.

    Google Scholar 

  • Finn BE, Forsén S. 1995 The evolving model of calmodulin structure, function and activation. Structure 3, 7–11.

    Google Scholar 

  • Flaherty KM, Zozulys S, Stryer L, McKay DB. 1993 Three dimensional structure of recoverin, a calcium sensor in vision. Cell 75, 709–716.

    Google Scholar 

  • Gellatly KS, Lefebvre DD 1993 Identification of a cDNA clone coding for a novel calcium-binding protein from potato tuber. Plant Physiol. 101, 1405–1406.

    Google Scholar 

  • Gilbert W. 1987 The exon theory of genes. Cold Spring Harbor Symp. Quant. Biol. 52, 901–905.

    Google Scholar 

  • Griffith JP, Kim JL, Kim EE, Sintchak MD, Thomson JA, Fitzgibbon MJ, Fleming MA, Caron PR, Hsiao K, Na via MA. X-ray structure of calcineurin inhibited by the immunophilin-immunosuppressant FKBP12-FK506 complex. 1995 Cell 82, 507–522.

    Google Scholar 

  • Grobler JA, Hurley JH. 1998 Catalysis by phospholipase Cd requires that Ca2+ bind to the catalytic domain, but not the C2 domain. Biochemistry 37, 5020–5028.

    Google Scholar 

  • Hardin PE, Angerer LM, Hardin SH, Angerer RC, Klein WH. 1988 Spec2 genes of Strongylocentrotus purpuratus. Structure and differential expression in embryonic aboral ectoderm cells. J. Mol. Biol. 202, 417–431.

    Google Scholar 

  • Head JF. 1989 Amino acid sequence of a low molecular weight, high affinity calcium-binding protein from the optic lobe of the squid, Loligo pealei. J. Biol. Chem. 264, 7202–7209.

    Google Scholar 

  • Heierhorst J, Kobe B, Feil SC, Parker MW, Benian GM, Weiss KR, Kemp BE. 1996 Ca2+/S100 regulation of giant protein kinases. Nature 380, 636–639.

    Google Scholar 

  • Hohenester E, Mauer P, Hohenadl D, Timpl R, Jansonius JN, Engel J. 1996 Structure of a novel extracellular Ca2+-binding module in BM-40. Nature Struct. Biol. 3, 67–73.

    Google Scholar 

  • Houdusse A, Cohen C. 1996 Structure of the regulatory domain of scallop myosin at 2 Å resolution: Implications for regulation. Structure 4, 21–32.

    Google Scholar 

  • Kawasaki H, Kretsinger RH. 1995 Calcium-binding proteins 1: EF-hands. Protein Profile 2, 305–490.

    Google Scholar 

  • Kissinger CR, Parge HE, Knighton DR, Lewis CT, Pelletier LA, Tempczyk A, Kalish VJ, Tucker KD, Showalter RE, Moomaw EW, Gastinel LN, Hakuba N, Chen X, Maldonado F, Barker JE, Bacquet R, Villafranca JE. 1995 Crystal structure of human calcineurin and the human FKBP12-FK506-calcineurin complex. Nature 378, 641–644.

    Google Scholar 

  • Kretsinger RH. 1972 Gene triplication deduced from the tertiary structure of a muscle calcium binding protein. Nature New Biology 240, 85–88.

    Google Scholar 

  • Kumar S, Harrylock M, Walsh KA, Cormier MJ, Charbonneau H. 1990 Amino acid sequence of the Ca2+-triggered luciferin binding protein of Renilla reniformis. FEBS Lett. 268, 287–290.

    Google Scholar 

  • Laroche A, Lemieux G, Pallotta D. 1989 The nucleotide sequence of a developmentally regulated cDNA from Physarum polysephalum. Nucleic Acids Res. 17, 10502–10502.

    Google Scholar 

  • Lee MG-S, Chen J, Ho AWM, D'Alesandro PA, Van der Ploeg LHT. 1990 A putative flagellar Ca2+-binding protein of trypanosomatid protozoan parasites. Nucleic Acids Res. 18, 4252–4252.

    Google Scholar 

  • Lee S-C, Kim I-G, Marekov LN, O'Keefe EJ, Parry DAD, Steinert PM. 1993 The structure of human trichohyalin. Potential multiple roles as a functional EF-hand-like calcium-binding protein, a cornified cell envelope precursor, and an intermediate filament-associated (cross-linking) protein. J. Biol. Chem. 268, 12164–12176.

    Google Scholar 

  • Lin CS, Shen W, Chen ZP, Tu YH, Matsudaira P. 1994 Identification of I-plastin, a human fimbrin isoform expressed in intestine and kidney. Mol. Cell. Biol. 14, 2457–2467.

    Google Scholar 

  • Lin X, Barber DL. 1996 A calcineurin homologous protein inhibits GTPase-stimulated Na-H exchange. Proc. Natl. Acad. Sci. 93, 12631–12636.

    Google Scholar 

  • Ling V, Zielinski RE. 1993 Isolation of an Arabidopsis cDNA sequence encoding a 22 kDa calcium-binding protein (CaBP-22) related to calmodulin. Plant Mol. Biol. 22, 207–214.

    Google Scholar 

  • Linse S, Thulin E, Gifford LK, Radzewsky D, Hagan J, Wilk RR, (kerfeldt KS. 1997 Domain organization of calbindin D28K as determined from the association of six synthetic EF-hand fragments. Prot. Sci. 6, 2385–2396.

    Google Scholar 

  • Lokuta AJ, Meyers MB, Sander PR, Fishman GI, Valdivia HH. 1997 Modulation of cardiac ryanodine receptors by sorcin. J. Biol. Chem. 272, 25333–25338.

    Google Scholar 

  • Lundberg S, Buevich AV, Sethson I, Edlund U, Backman L. 1997 Calcium-binding mechanism of human nonery-throid α-spectrin EF-structures. Biochemistry 23, 7199–7208.

    Google Scholar 

  • Markova NG, Marekov LN, Chipev CC, Gan S-Q, Idler WW, Steinert PM. 1993 Profilaggrin is a major epidermal calcium-binding protein. Mol. Cell. Biol. 13, 613–625.

    Google Scholar 

  • Nakayama S, Kretsinger RH. 1994 Evolution of the EF-hand family of proteins. Annu. Rev. Biophys. Biomol. Structure 23, 473–507.

    Google Scholar 

  • Orengo CA, Jones, DT, Thornton JM. 1994 Protein super-families and domain superfolds. Nature 372, 631–634.

    Google Scholar 

  • Ozawa M, Muramatsu T. 1993 Reticulocalbin, a novel endoplasmic reticulum resident Ca2+-binding protein with multiple EF-hand motifs and a carboxyl-terminal HDEL sequence. J. Biol. Chem. 268, 699–705.

    Google Scholar 

  • Prasher DC, McCann RO, Longiaru M, Cormier MJ. 1987 Sequence comparisons of complementary DNAs encoding aequorin isotypes. Biochemistry 26, 1326–1332

    Google Scholar 

  • Rawlings DJ, Kaslow DC. 1992 A novel 40-kDa membrane-associated EF-Hand calcium-binding protein in Plasmodium falciparum. J. Biol. Chem. 267 3976–3982.

    Google Scholar 

  • Rayment I, Rypniewski WR, Schmidt-Bäse K, Smith R, Tomchick DR, Benning MM, Winkelmann DA, Wesenberg G, Holden HM. 1993 Three-dimensional structure of myosin subfragment-1: A molecular motor. Science 261, 50–58.

    Google Scholar 

  • Rhoads A, Friedberg F. 1997 Sequence motifs for calmodulin recognition. FASEB J. 11, 331–340.

    Google Scholar 

  • Rustandi RR, Drohat AC, Weber DJ. 1998 The Ca2+-dependent interaction of S100B(ββ) with a peptide derived from p53. Biochemistry 37, 1951–1960.

    Google Scholar 

  • Saijo Y, Hata S, Sheen J, Izui K. 1997 cDNA cloning and prokaryotic expression of maize calcium-dependent protein kinases. Biochim. Biophys. Acta 1350, 109–114.

    Google Scholar 

  • Salvato M, Sulston J, Albertson D, Brenner S. 1986 A novel calmodulin-like gene from the nematode Caenorhabditis elegans. J. Mol. Biol. 190, 281–289.

    Google Scholar 

  • Sauter A, Staudenmann W, Hughes GJ, Heizmann, CW. 1995 A novel EF-hand Ca2+-binding protein from abdominal muscle of crustaceans with similarity to calcyphosine from dog thyrodea. Eur. J. Biochem 227, 97–101.

    Google Scholar 

  • Schäfer BW, Wicki R, Engelkamp D, Mattei M-G, Heizmann CW. 1995 Isolation of a YAC clone covering a cluster of nine S100 genes on human chromosome 1q21: Rationale for a new nomenclature of the S100 calcium-binding protein family. Genomics 25, 638–643.

    Google Scholar 

  • Schäfer BW, Heizmann CW. 1996 The S100 family of EF-hand calcium-binding proteins: functions and pathology. TIBS 21, 134–140.

    Google Scholar 

  • Smith SP, Shaw GS. 1998 A novel calcium-sensitive switch revealed by the structure of human S100B in the calcium-bound form. Structure 6, 211 222.

    Google Scholar 

  • Steele FR, Washburn T, Rieger R, O'Tousa JE. 1992 Drosophila retinal degeneration C (rdgC) encodes a novel serine/threonine protein phosphatase. Cell 69, 669–676.

    Google Scholar 

  • Swan DG, Cortes J, Hale RS, Leadlay PF. 1989 Cloning, characterization, and heterologous expression of the Saccharopolyspora eythraea (Streptomyces erythraeus) gene encoding an EF-Hand calcium-binding protein. J Bacteriol. 171, 5614–5619.

    Google Scholar 

  • Tachikui H, Navet AF, Ozawa M. 1997 Identification of the Ca2+-binding domains in reticulocalbin, an endoplasmic reticulum resident Car2+-binding protein with multiple EF-hand motifs. J. Biochem. (Tokyo) 121, 145–149.

    Google Scholar 

  • Takemasa T, Takagi T. Kobayashi T, Konishi K, Watanabe Y. 1990 The third calmodulin family protein in Tetrahymena. Cloning of the cDNA for Tetrahymena calcium-binding protein of 23 kDa (TCBP-23). J. Biol. Chem. 265, 2514–2517.

    Google Scholar 

  • Taylor KA, Taylor DW. 1993 Projection image of smooth muscle α-actinin from 2-D crystals formed on positively charged lipid layers J. Mol. Biol. 230, 196–205.

    Google Scholar 

  • Templeton TJ, Fujioka H, Aikawa M, Parker KC. 1997 Plasmodium falciparum Pfs40, renamed Pf39, is localized to an intracellular membrane-bound compartment and is not sexual stage-specific. Mol. Biochem. Parasitol. 90, 359–365.

    Google Scholar 

  • Vassylev DG, Takeda S, Wakatsuki S, Maeda K, Maeda Y. 1998 Crystal structure of troponin C in complex with troponin I fragment at 2.3 Å resolution. Proc. Natl. Acad. Sci. 95 4847–4852.

    Google Scholar 

  • Wong S, Morales TH, Neigel JE, Campbell DA. 1993 Genomic and transcriptional control linkage of the genes for calmodulin, EF-hand 5 protein, and ubiquitin extension protein 52 in Trypanosoma brucei. Mol. Cell. Biol. 13, 207–216.

    Google Scholar 

  • Wooding FBP, Morgan G, Jones GV, Care AD. 1996 Calcium transport and the localisation of calbindin D9k in the ruminant placenta during the second half of pregnancy. Cell Tissue Res. 285, 477–489.

    Google Scholar 

  • Wu Y, Haghighat NG, Ruben L. 1992 The predominant calcimedins from Trypanosoma brucei comprise a family of flagellar EF-hand calcium-binding proteins. Biochem. J. 287, 187–193.

    Google Scholar 

  • Xiang M, Ge T, Tomlinson CR, Klein WH. 1991 Structure and promoter activity of the LpS1 genes of Lytechinus pictus. J. Biol. Chem. 266, 10524–10533.

    Google Scholar 

  • Yamada K, Sakane F, Matsushima N, Kanoh H. 1997 EF-hand motifs of α, β and γ isoforms of diacylglycerol kinase bind calcium with different affinities and conformational changes. Biochem J. 321, 59–64.

    Google Scholar 

  • Zhao Y, Kappes B, Franklin RM. 1993 Gene structure and expression of an unusual protein kinase from Plasmodium falciparum homologous at its carboxyl terminus with the EF Hand calcium-binding proteins. J. Biol. Chem. 268, 4347–4354.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kawasaki, H., Nakayama, S. & Kretsinger, R.H. Classification and evolution of EF-hand proteins. Biometals 11, 277–295 (1998). https://doi.org/10.1023/A:1009282307967

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009282307967

Navigation