Cloning and characterization of the yak gene coding for calpastatin and in silico analysis of its putative product
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The calcium-activated neutral proteases, μ- and m-calpain, along with their inhibitor, calpastatin, have been demonstrated to mediate a variety of Ca2+-dependent processes including signal transduction, cell proliferation, cell cycle progression, differentiation, apoptosis, membrane fusion, platelet activation and skeletal muscle protein degradation. The cDNA coding for yak calpastatin was amplified and cloned by RT-PCR to investigate and characterize the nucleotide/amino-acid sequence and to predict structure and function of the calpastatin. The present study suggests that the yak calpastatin gene encodes a protein of 786 amino acids that shares 99 % sequence identity with the amino-acid sequence of cattle calpastatin, and that the yak protein is composed of an N-terminal region (domains L and XL) and four repetitive homologous C-terminal domains (d1–d4), in which several prosite motifs are present including short peptide L54–64 (EVKPKEHTEPK in domain L) and GXXE/ DXTIPPXYR (in subdomain B), where X is a variable amino acid. Our results suggest the existence of other functional sites including potential phosphorylation sites for protein kinase C, cAMP- and cGMP-dependent protein kinase, casein kinase II, as well as N-myristoylation and amidation sites that play an important role in molecular regulation of the calpain/calpastatin system. The regulation of the calpain/calpastatin system is determined by the interaction between dIV and dVI in calpains and subdomains A, B, and C in calpastatin.
- Bendtsen JD, Nielsen H, Heijne G, Brunak S (2004) Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340: 783-795.
- Betts R, Weinsheimer S, Blouse GE, Anagli J (2003) Structural determinants of the calpain inhibitory activity of calpastatin peptide B27-WT. J Biol Chem 278: 7800-7809.
- Carragher NO, Westhoff MA, Riley D, Potter DA, Dutt P, Elce JS, Greer PA, Frame MC (2002) v-Src-induced modulation of the calpain-calpastatin proteolytic system regulates transformation. Mol Cell Biol 22: 257-269.
- Chou PY (1990) Prediction of protein structural classes from amino acid composition. In Prediction of protein structure and the principles of protein conformation. Plenum Press, New York, NY.
- Chou PY, Fasman GD (1978) Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol 47: 45-148.
- Croall DE, Demartino GN (1991) Calcium-activated neutral protease (calpain) system: structure, function, and regulation. Physiol Rev 71: 813-847.
- Emori Y, Kawasaki H, Imajoh S, Minami Y, Suzuki K (1988) All four repeating domains of the endogenous inhibitor for calcium-dependent protease independently retain inhibitory activity. J Biol Chem 263: 2364-2370.
- Glading A, Lauffenburger DA, Wells A (2002) Cutting to the chase: calpain proteases in cell motility. Trends Cell Biol 12: 46-54.
- Goll DE, Thompson VF, Li H, Wei W, Cong J (2003) The calpain system. Physiol Rev 83: 731-801.
- Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41: 95-98.
- Hanna RA, Robert LC, Peter LD (2008) Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin. Nature 456: 409-412.
- Hanna RA, Garcia-Diaz BE, Davies PL (2007) Calpastatin simultaneously binds four calpains with different kinetic constants. FEBS Lett 581: 2894-2898.
- Hao LY, Kameyama A, Kuroki S, Takano J, Takano E, Maki M, Kameyama M (2000) Calpastatin domain L is involved in the regulation of L-type Ca2+ channels in guinea pig cardiac myocytes. Biochem Biophys Res Commun 279: 756-761.
- Hirose K, Kadowaki S, Tanabe M, Takeshima H, Iino M (1999) Spatiotemporal dynamics of inositol 1,4,5-trisphosphate that underlies complex Ca2+ mobilization patterns. Science 284: 1527-1530.
- Kawasaki H, Emori Y, Imajoh-ohmi S, Minami Y, Suzuki K (1989) Identification and characterization of inhibitory sequences in four repeating domains of the endogenous inhibitor for calcium dependent protease. J Biochem 106: 274-281.
- Kiss R, Bozoky Z, Kovacs D, Rona G, Friedrich P, Dvortsak P, Weisemann R, Tompa P, Perczel A (2008a) Calcium-induced tripartite binding of intrinsically disordered calpastatin to its cognate enzyme, calpain. FEBS Lett 582: 2149-2154.
- Kiss R, Kovacs D, Tompa P, Perczel A (2008b) Local structural preferences of calpastatin, the intrinsically unstructured protein inhibitor of calpain. Biochemistry 47: 6936-6945.
- Maki M, Takano E, Mori H, Sato A, Murachi T, Hatanaka M (1987) All four internally repetitive domains of pig calpastatin possess inhibitory activities against calpains I and II. FEBS Lett 223: 174-180.
- Maki M, Takano E, Osawa T, Ooi T, Murachi T, Hatanaka M (1988) Analysis of structure-function relationship of pig calpastatin by expression of mutated cDNAs in Escherichia coli. J Biol Chem 263: 10254-10261.
- Melloni E, Averna M, Stifanese R, Tullio RD, Defranchi E, Salamino F, Pontremoli S (2006) Association of calpastatin with inactive calpain. J Biol Chem 281: 24945-24954.
- Minobe E, Hao LY, Saud ZA, Xu JJ, Kameyama A, Maki M, Jewell KK, Parr T, Bardsley RG, Kameyama M (2006) A region of calpastatin domain L that reprimes cardiac L-type Ca2+ channels. Biochem Biophys Res Commun 348: 288-294.
- Moldoveanu T, Gehring K, Green DR (2008) Concerted multi-pronged attack by calpastatin to occlude the catalytic cleft of heterodimeric calpains. Nature 456: 404-408.
- Mucsi Z, Hudecz F, Hollósi M, Tompa P, Friedrich P (2003) Binding-induced folding transitions in calpastatin subdomains A and C. Protein Sci 12: 2327-2336.
- Pfizer J, Assfalg-Machleidt I, Machleidt W, chaschke N (2008) Inhibition of human mu-calpain by conformationally constrained calpastatin peptides. Biol Chem 389: 83-90.
- Sambrook J, Russell D (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Press, Cold Spring Harbor, NY.
- Shiraha H, Glading A, Chou J, Jia Z, Wells A (2002) Activation of m-calpain (calpain II) by epidermal growth factor is limited by protein kinase A phosphorylation of m-calpain. Mol Cell Biol 22: 2716-2727.
- Takano J, Watanabe M, Hitomi K, Maki M (2000) Four types of calpastatin isoforms with distinct amino-terminal sequences are specified by alternative first exons and differentially expressed in mouse tissues. J Biochem (Tokyo) 128: 83-92.
- Todd B, Moore D, Deivanayagam CC, Lin GD, Chattopadhyay D, Maki M, Wang KK, Narayana SV (2003) A structural model for the inhibition of calpain by calpastatin: crystal structures of the native domain VI of calpain and its complexes with calpastatin peptide and a small molecule inhibitor. J Mol Biol 328: 131-146.
- Toke O, Banoczi Z, Tarkanyi G, Friedrich P, Hudecz F (2009) Folding transitions in calpain activator peptides studied by solution NMR spectroscopy. J Pept Sci 15: 404-410.
- Tompa P, Mucsi Z, Orosz G, Friedrich P (2002) Calpastatin subdomains A and C are activators of calpain. J Biol Chem 277: 9022-9026.
- Tullio RD, Averna M, Salamino F, Pontremoli S, Melloni E (2000) Differential degradation of calpastatin by μ- and m-calpain in Ca2+-enriched human neuroblastoma LAN-5 cells. FEBS Lett 475: 17-21.
- Uemori T, Shimojo T, Asada K, Asano T, Kimizuka F, Kato I, Maki M, Hatanaka M, Murachi T, Hanzawa H, Arata Y (1990) Characterization of a functional domain of human calpastatin. Biochem Biophys Res Commun 166: 1485-1493.
- Weber H, Jonas L, Hühns S, Schuff-Werner P (2004) Dysregulation of the calpain-calpastatin system plays a role in the development of cerulein-induced acute pancreatitis in the rat. Am J Physiol Gastrointest Liver Physiol 286: G932-G941.
- Wendt A, Thompson VF, Goll DE (2004) Interaction of calpastatin with calpain: a review. Biol Chem 385: 465-472.
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