Abstract
Calcium-activated neutral protease (CANP) is a typical intracellular protease responsible for the processing and turnover of various intracellular proteins.1–5 Uncontrolled, CANP can destroy various cellular components, leading to various disease processes like muscular dystrophy. Hence the activity must be tightly regulated. Because CANP absolutely requires calcium for activity, calcium plays a pivotal role in its regulation. Although many of the important steps of CANP activity regulation have been clarified, most studies have not provided any insight into the binding of calcium to CANP6. Structural analyses of CANP have revealed the existence of a calmodulin-like domain in the C-terminal region of each of the two constituent, subunits (80K and 30K subunits).7–9 These domains are presumed to bind calcium and determine the calcium sensitivity of CANP. Nevertheless, the following questions remain unsolved: whether the calmodulin domains indeed bind calcium; whether calcium-binding sites exist only in the calmodulin domains; and, if so, how many calcium ions bind to each domain and which EF hand structures in the calmodulin domains are functional calcium-binding sites.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
K. Suzuki, Calcium-activated neutral protease: domain structure and activity regulation, Trend. Biol. Sci.’ 12:103 (1987).
S. Pontremoli and E. Melloni, Extralysosomal degradation, Ann. Rev. Biochem., 55:455 (1986).
K. Suzuki, S. Ohno, Y. Emori, S. Imajoh, and H. Kawasaki, Calcium-activated neutral protease (CANP) and its biological and medical implications, Prog. Clin. Biochem. Med., 5:43 (1987).
T. Murachi, M. Hatanaka, and T. Hamakubo, Calpains and neuropeptide metabolism, in “Neuropeptides and their peptidases,” Turner, ed., Ellis Horwood Ltd., Chichester (1987).
R. Mellgren, Calcium-dependent proteases: an enzyme system active at cellular membrane? FASEB J., 2:110 (1988).
K. Suzuki and S. Ishiura, Effect of metal ions on the activity of calcium-activated neutral protease (CANP), J. Biochem., 93:1463 (1983).
S. Ohno, Y. Emori, S. Imajoh, H. Kawasaki, M. Kisaragi, and K. Suzuki, Evolutionary origin of calcium dependent protease by fusion of genes for a thiol protease and a calcium binding protein? Nature, 312:566 (1984).
Y. Emori, H. Kawasaki, S. Imajoh, S. Kawashima, and K. Suzuki, Isolation and sequence analysis of cDNA clones for the small subunit of rabbit calcium-dependent protease, J. Biol. Chem., 261:9472 (1986).
T. Sakihama, H. Kakidani, K. Zenita, N. Yumoto, T. Kikuchi, T. Sasaki, R. Knnagi, S. Nakanishi, M. Ohmori, K. Takio, K. Titani, and T. Murachi, A putative Ca2+-binding protein: structure of the light subunit of porcine calpain elucidated by molecular cloning and protein sequence analysis, Proc. Natl. Acad. Sci.USA, 82:6075 (1985).
H. Kawasaki, H. Kasai, and T. Okuyama, Protein analyses and reagents: microscale assay of calcium-binding activity of proteins and peptides using a nitrocellulose membrane, Anal. Biochem., 148:297 (1985).
K. Maruyama, T. Mikawa, and S. Ebashi, Detection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecylsulfate gel electrophoresis, J. Biochem., 95:511 (1984).
Y. Emori, H. Kawasaki, H. Sugihara, S. Imajoh, S. Kawashima, and K. Suzuki, Isolation and sequence analyses of cDNA clones for the large subunits of two isozymes of rabbit calcium-dependent protease, J. Biol. Chem., 261:9472 (1986).
Y. Minami, Y. Emori, H. Kawasaki, and K. Suzuki, E-F hand structure domain of calcium-activated neutral protease (CANP) can bind Ca ions, J. Biochem., 101:889 (1987).
Y. Minami, Y. Emori, S. Imajoh, H. Kawasaki, and K. Suzuki, Carboxy-terminal truncation and site-directed mutagenesis of the small subunit of rabbit calcium-dependent protease, J. Biochem., 103: in press (1988).
S. Imajoh, H. Kawasaki, and K. Suzuki, The COOH-terminal EF hand structure of calcium activated neutral protease (CANP) is important for the association of subunits and resulting protease activity, J. Biochem., 101:447 (1987).
T. Kikuchi, N. Yumoto, T. Sasaki, and T. Murachi, Reconstitution of calpain I and calpain II from their subunits. Interchangeability of the light subunits, Arch. Biophys. Biochem., 234:639 (1984).
O.H. Lowry, N.I. Rosenbrough, A.L. Farr, and R.J. Randall, Protein measurement with the Folin-phenol reagent, J. Biol. Chem., 193: 265 (1955).
U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227:680 (1970).
H. Kawasaki, S. Imajoh, S. Kawashima, H. Hayashi, and K. Suzuki, The small subunit of calcium dependent protease with different calcium sensitivities are identical, J. Biochem., 99:1525 (1986).
F.C. Reinach, K. Nagai, and J. Kendrick-Jones, Site-directed mutagenesis of the regulatory light chain Ca2+ /Mg2+ binding site and its role in hybrid myosins, Nature, 322:80 (1986).
R.M. Tufty, R.H. Kretsinger, Troponin and parvalbumin calcium binding regions predicted in myosin light chain and T4 lysozyme, Science, 187:167 (1975).
S. Imajoh, H. Kawasaki, and K. Suzuki, Limited autolysis of calcium-activated neutral protease (CANP): reduction of the Ca2+-requirement is due to the N-terminal processing of the large subunit, J. Biochem., 100:633 (1986).
Y. Emori, S. Ohno, M. Tobita, and K. Suzuki, Gene structure of calcium-dependent protease retains the ancestral organization of the calcium-binding protein gene, FEBS Letters, 194:249 (1986).
S. Miyake, Y. Emori, and K. Suzuki, Gene organization of the small subunit of human calcium-activated neutral protease, Nucleic Acids Res., 14:8805 (1986).
K. Suzuki, S. Imajoh, Y. Emori, H. Kawasaki, Y. Minami, and S. Ohno, Calcium-activated neutral protease and its endogenous inhibitor. Activationat the cell membrane and biological function, FEBS Letters, 220:271 (1987).
K. Suzuki, S. Imajoh, Y. Emori, H. Kawasaki, Y. Minami, and S. Ohno, Regulation of activity of calcium-activated neutral protease, Adv. Enzyme Regul., 27:153 (1988).
K. Suzuki, S. Tsuji, K. Kubota, Y. Kimura, and K. Suzuki, Limited autolysis of Ca2+-activated neutral protease changes its sensitivity to Ca2+ ions, J. Biochem., 90:275 (1981).
K. Suzuki, Reaction of calcium-activated neutral protease (CANP) with an epoxysuccinyl derivative (E64c) and iodoacetic acid, J. Biochem., 93: 1305 (1983).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1989 Plenum Press, New York
About this chapter
Cite this chapter
Suzuki, K., Minami, Y., Emori, Y., Imajoh, S., Kawasaki, H. (1989). Analysis of Calcium-Binding Sites in Calcium-Activated Neutral Protease. In: Hidaka, H., Carafoli, E., Means, A.R., Tanaka, T. (eds) Calcium Protein Signaling. Advances in Experimental Medicine and Biology, vol 255. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5679-0_19
Download citation
DOI: https://doi.org/10.1007/978-1-4684-5679-0_19
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-5681-3
Online ISBN: 978-1-4684-5679-0
eBook Packages: Springer Book Archive