Overexpression, purification, and characterization of human m-calpain and its active site mutant, m-C105S-calpain, using a baculovirus expression system

Journal of Biochemistry

Volumn 124, Issue 5, 1998, Pages 957-961

  Masumoto, Hajime ^a   Yoshizawa, Toshio ^a   Sorimachi, Hiroyuki ^a   Nishino, Takeshi ^b   Ishiura, Shoichi ^a   Suzuki, Koichi ^a  

^a UNIVERSITY OF TOKYO   (Japan)

^b NIPPON MEDICAL SCHOOL   (Japan)

Author keywords

Baculovirus; M calpain; Mutant; Overexpression; Sf 9 cell

Indexed keywords

2 DIETHYLAMINOETHANOL; CALCIUM ION; CALPAIN; CYSTEINE; MUTANT PROTEIN; RECOMBINANT ENZYME; SERINE;

ANIMAL TISSUE; ARTICLE; AUTOLYSIS; BACULOVIRUS; COLUMN CHROMATOGRAPHY; ENZYME ACTIVE SITE; ENZYME ACTIVITY; ENZYME SUBSTRATE; ENZYME SUBUNIT; GEL FILTRATION; GENE EXPRESSION SYSTEM; GENE OVEREXPRESSION; HUMAN; NONHUMAN; PURIFICATION; RABBIT; SITE DIRECTED MUTAGENESIS;

ANIMALIA; ORYCTOLAGUS CUNICULUS; UNIDENTIFIED BACULOVIRUS;

EID: 0031755539     PISSN: 0021924X     EISSN: None     Source Type: Journal    
DOI: 10.1093/oxfordjournals.jbchem.a022213     Document Type: Article

References (24)

1. Mellgren, R.L. (1987) Calcium-dependent proteases: an enzyme system active at cellular membranes? FASEB J. 1, 110-115
2. Wang, K.K.W., Villalobo, A., and Roufogalis, B.D. (1989) Calmodulin-binding proteins as calpain substrates. Biochem. J. 262, 693-706
3. Suzuki, K., Sorimachi, H., Yoshizawa, T., Kinbara, K., and Ishiura, S. (1995) Calpain: Novel family members activation, and physiological function. Biol. Chem. Hoppe-Seyler 376, 523-529
4. Johnson, P. (1990) Calpains (intracellular calcium-activated cysteine proteinases): Structure-activity relationships and involvement in normal and abnormal cellular metabolism. Int. J. Biochem. 22, 811-822
5. Sorimachi, H., Ishiura, S., and Suzuki, K. (1997) Structure and physiological function of calpains. Biochem. J. 328, 721-732
6. Graham-Siegenthaler, K., Gauthier, S., Davies, P.L., and Elce, J.S. (1994) Active recombinant rat calpain II. J. Biol. Chem. 269, 30457-30460
7. Meyer, S.L., Bozyczko-Coyne, D., Mallya, S.K., Spais, C.M., Bihovsky, R., Kawooya, J.K., Lang, D.M., Scott, R.W., and Siman, R. (1996) Biologically active monomeric and heterodimeric recombinant human calpain I produced using the baculovirus expression system. Biochem. J. 314, 511-519
8. Sorimachi, H., Imajoh-Ohmi, S., Emori, Y., Kawasaki, H., Ohno, S., Minami, Y., and Suzuki, K. (1989) Molecular cloning of a novel mammalian calcium-dependent protease distinct from both m-and μ-types. J. Biol. Chem. 264, 20106-20111
9. 2+-binding domain. J. Biol. Chem. 268, 19476-19482
10. Inomata, M., Nomoto, M., Hayashi, M., Nakamura, M., Imahori, K., and Kashima, S. (1984) Comparison of low and high calcium requiring forms of the calcium-activated neutral protease (CANP) from rabbit skeletal muscle. J. Biochem. 95, 1661-1670
11. 2+-activated neutral protease. Biochemistry 27, 8122-8128
12. Kunkel, T.A. (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. USA 82, 488-492
13. Ohno, S., Emori, Y., and Suzuki, K. (1986) Nucleotide sequence of a cDNA coding for the small subunit of human calcium-dependent protease. Nucleic Acids Res. 14, 5559
14. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254
15. 2-terminal processing of the large subunit. J. Biochem. 100, 633-642
16. Sasaki, T., Kikuchi, T., Yumoto, N., Yoshimura, N., and Murachi, T. (1984) Comparative specificity and kinetic studies on porcine calpain I and calpain II with naturally occurring peptides and synthetic fluorogenic substrates. J. Biol. Chem. 259, 12489-12494
17. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685
18. Yoshizawa, T., Sorimachi, H., Tomioka, S., Ishiura, S., and Suzuki, K. (1995) A catalytic subunit of calpain possesses full proteolytic activity. FEBS Lett. 358, 101-103
19. 2+-activated neutral protejnase of rabbit skeletal muscle. J. The characterization of the 80K and the 30K subunits. J. Biochem. 90, 233-240
20. Suzuki, K., Ishiura, S., Tsuji, S., Katamoto, T., Sugita, H., and Imahori, K. (1979) Calcium activated neutral protease from human skeletal muscle. FEBS Lett. 104, 355-358
21. Clark, P.I. and Lowe, G. (1978) Conversion of the active-site cysteine residue of papain into a dehydro-serine, a serine and a glycine residue. Eur. J. Biochem. 84, 293-299
22. Urade, R., Oda, T., Ito, H., Moriyama, T., Utsumi, S., and Kito, M. (1997) Functions of characteristic Cys-Gly-His-Cys (CGHC) and Gln-Glu-Asp-Leu (QEDL) motifs of microsomal ER-60 protease. J. Biochem. 122, 834-842
23. 2+-induced conformational changes. Nat. Struct. Biol. 4, 532-538
24. Lin, G., Chattopadhyay, D., Maki, M., Wang, K.K.W., Carson, M., Jin, L., Yuen, P., Takano, E., Hatanaka, M., DeLucas, L.J., and Narayana, S.V.L. (1997) Crystal structure of calcium bound domain VI of calpain at 1.9 Å resolution and its role in enzyme assembly, regulation, and inhibitor binding. Nat. Struct. Biol. 4, 539-547