HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Emori, Y. Articles by Suzuki, K. Search for Related Content PubMed PubMed Citation Articles by Emori, Y. Articles by Suzuki, K. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 263, Issue 5, 2364-2370, Feb, 1988

All four repeating domains of the endogenous inhibitor for calcium- dependent protease independently retain inhibitory activity. Expression of the cDNA fragments in Escherichia coli

Y Emori, H Kawasaki, S Imajoh, Y Minami and K Suzuki
Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Japan.

We have already determined the primary structure of the endogenous inhibitor for calcium-dependent protease (CANP inhibitor, calpastatin) from the cDNA sequence and revealed that the CANP inhibitor contains four internally repeating units which could be responsible for its multiple reactive sites (Emori, Y., Kawasaki, H., Imajoh, S., Imahori, K., and Suzuki, K. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 3590-3594). Restriction fragments of the cDNA corresponding to each of the four domains (encoding 104-156 amino acid residues of the total 718 residues) were subcloned into the multicloning site of pUC9 or pUC18 in a direction and frame matched to the lacZ' open reading frame of the vector. Under the lac operator-promoter system, we succeeded in producing truncated fragments of the CANP inhibitor in Escherichia coli. The CANP inhibitor fragments were partially purified, and the inhibitory activities toward calcium-dependent protease (CANP) were examined. All fragments containing well conserved regions of about 30 amino acid residues (domains I-IV) located in the middle of the four units exhibited the inhibitory activity. However, their inhibitory activities varied considerably. Further truncation experiments revealed that small fragments containing 30-70 amino acid residues of the CANP inhibitor still retained inhibitory activity. From these experimental results the following conclusions can be drawn: 1) each of the four repeating units of the CANP inhibitor (about 140 amino acid residues) is a real functional unit and can inhibit CANP activity independently; and 2) domains corresponding to well conserved sequences of about 30 amino acid residues containing a consensus Thr-Ile-Pro-Pro-X-Tyr-Arg sequence are essential for the inhibitory activity, and the bordering regions are important for its modulation.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				E. Melloni, M. Averna, R. Stifanese, R. De Tullio, E. Defranchi, F. Salamino, and S. Pontremoli Association of Calpastatin with Inactive Calpain: A NOVEL MECHANISM TO CONTROL THE ACTIVATION OF THE PROTEASE? J. Biol. Chem., August 25, 2006; 281(34): 24945 - 24954. [Abstract] [Full Text] [PDF]

				M. Bokor, V. Csizmok, D. Kovacs, P. Banki, P. Friedrich, P. Tompa, and K. Tompa NMR Relaxation Studies on the Hydrate Layer of Intrinsically Unstructured Proteins Biophys. J., March 1, 2005; 88(3): 2030 - 2037. [Abstract] [Full Text] [PDF]

				P. Tompa, P. Buzder-Lantos, A. Tantos, A. Farkas, A. Szilagyi, Z. Banoczi, F. Hudecz, and P. Friedrich On the Sequential Determinants of Calpain Cleavage J. Biol. Chem., May 14, 2004; 279(20): 20775 - 20785. [Abstract] [Full Text] [PDF]

				Z. Mucsi, F. Hudecz, M. Hollosi, P. Tompa, and P. Friedrich Binding-induced folding transitions in calpastatin subdomains A and C Protein Sci., October 1, 2003; 12(10): 2327 - 2336. [Abstract] [Full Text] [PDF]

				D. E. GOLL, V. F. THOMPSON, H. LI, W. WEI, and J. CONG The Calpain System Physiol Rev, July 1, 2003; 83(3): 731 - 801. [Abstract] [Full Text] [PDF]

				C. Fukiage, E. Nakajima, H. Ma, M. Azuma, and T. R. Shearer Characterization and Regulation of Lens-specific Calpain Lp82 J. Biol. Chem., May 31, 2002; 277(23): 20678 - 20685. [Abstract] [Full Text] [PDF]

				P. Tompa, Z. Mucsi, G. Orosz, and P. Friedrich Calpastatin Subdomains A and C Are Activators of Calpain J. Biol. Chem., March 8, 2002; 277(11): 9022 - 9026. [Abstract] [Full Text] [PDF]

				J. Huang and N. E. Forsberg Role of calpain in skeletal-muscle protein degradation PNAS, October 13, 1998; 95(21): 12100 - 12105. [Abstract] [Full Text] [PDF]

				M. Cong, V. F. Thompson, D. E. Goll, and P. B. Antin The Bovine Calpastatin Gene Promoter and a New N-terminal Region of the Protein Are Targets for cAMP-dependent Protein Kinase Activity J. Biol. Chem., January 2, 1998; 273(1): 660 - 666. [Abstract] [Full Text] [PDF]