IDENTIFICATION AND CHARACTERIZATION OF AN ACTIN-BINDING SITE OF CAPZ

被引:65
作者
HUG, C [1 ]
MILLER, TM [1 ]
TORRES, MA [1 ]
CASELLA, JF [1 ]
COOPER, JA [1 ]
机构
[1] JOHNS HOPKINS UNIV, SCH MED, DEPT PEDIAT, DIV HEMATOL, BALTIMORE, MD 21205 USA
关键词
D O I
10.1083/jcb.116.4.923
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
A mAb (1E5) that binds the COOH-terminal region of the beta-subunit of chicken CapZ inhibits the ability of CapZ to bind the barbed ends of actin filaments and nucleate actin polymerization. CapZ prepared as fusion proteins in bacteria or nonfusion proteins by in vitro translation has activity similar to that of CapZ purified from muscle. Deletion of the COOH-terminus of the beta-subunit of CapZ leads to a loss of CapZ's ability to bind the barbed ends of actin filaments. A peptide corresponding to the COOH-terminal region of CapZ beta, expressed as a fusion protein, binds actin monomers. The mAb 1E5 also inhibits the binding of this peptide to actin. These results suggest that the COOH-terminal region of the beta-subunit of CapZ is an actin-binding site. The primary structure of this region is not similar to that of potential actin-binding sites identified in other proteins. In addition, the primary structure of this region is not conserved across species.
引用
收藏
页码:923 / 931
页数:9
相关论文
共 26 条
[11]   EFFECT OF CAPPING PROTEIN ON THE KINETICS OF ACTIN POLYMERIZATION [J].
COOPER, JA ;
POLLARD, TD .
BIOCHEMISTRY, 1985, 24 (03) :793-799
[12]   VARIANT CDNAS ENCODING PROTEINS SIMILAR TO THE ALPHA-SUBUNIT OF CHICKEN CAPZ [J].
COOPER, JA ;
CALDWELL, JE ;
GATTERMEIR, DJ ;
TORRES, MA ;
AMATRUDA, JF ;
CASELLA, JF .
CELL MOTILITY AND THE CYTOSKELETON, 1991, 18 (03) :204-214
[13]  
EY PL, 1986, METHOD ENZYMOL, V121, P497
[14]  
HARLOW E, 1988, ANTIBODIES LABORATOR
[15]  
HARTMANN H, 1989, J BIOL CHEM, V264, P12639
[16]   REGULATION OF CAPZ, AN ACTIN CAPPING PROTEIN OF CHICKEN MUSCLE, BY ANIONIC PHOSPHOLIPIDS [J].
HEISS, SG ;
COOPER, JA .
BIOCHEMISTRY, 1991, 30 (36) :8753-8758
[17]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[18]  
POLLARD TD, 1982, METHOD ENZYMOL, V85, P211
[19]   ACTIN AND ACTIN-BINDING PROTEINS - A CRITICAL-EVALUATION OF MECHANISMS AND FUNCTIONS [J].
POLLARD, TD ;
COOPER, JA .
ANNUAL REVIEW OF BIOCHEMISTRY, 1986, 55 :987-1035
[20]  
SAFER D, 1991, J BIOL CHEM, V266, P4029