title: Proteolysis of acidic calponin by mu-calpain.
abstract: Acidic calponin is an actin binding protein expressed in smooth muscle and brain. Although the role of smooth muscle calponin (basic calponin) has been well studied, few studies have been performed on acidic calponin. In the present study, we demonstrated that acidic calponin binds to filamentous actin, but not monomeric actin. A co-sedimentation assay indicated that acidic calponin binds to actin with an apparent binding constant of 4 x 10(5) M(-1). In the presence of an excess amount of calmodulin, the binding of acidic calponin to actin was inhibited. The binding of acidic calponin to calmodulin was Ca(2+)-dependent with K(d) of 31 microM. We next investigated whether or not acidic calponin could be a substrate for mu-calpain in vitro, since it has been shown that basic calponin is cleaved by mu-calpain. The results showed that acidic calponin was also cleaved by mu-calpain. Neither the proteolytic pattern nor velocity of acidic calponin was different in the absence or presence of calmodulin. When acidic calponin had bound to actin, however, the susceptibility of the acidic calponin to mu-calpain was significantly reduced, which was reversed by the addition of calmodulin. Our results suggest that acidic calponin might be involved in the mu-calpain-regulated actin cytoskeleton.
authors.affiliation: Department of Veterinary Pharmacology, Graduate school of Agriculture and Life Sciences, The University of Tokyo, Yayoi Bunkyo-ku, Tokyo 113, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Yoshimoto
authors.forename: M
authors.initials: M
authors.lastname: Hori
authors.forename: H
authors.initials: H
authors.lastname: Ozaki
authors.forename: H
authors.initials: H
authors.lastname: Karaki
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008840:Microfilament Proteins; D011817:Rabbits; D013552:Swine; D000096985:Calponins
chemical_list: D002135:Calcium-Binding Proteins; D002154:Calpain; D008840:Microfilament Proteins; D000096985:Calponins