Characterization of gelsolin truncates that inhibit actin depolymerization by severing activity of gelsolin and cofilin

被引:13
作者
Fujita, H [1 ]
Allen, PG [1 ]
Janmey, PA [1 ]
Azuma, T [1 ]
Kwiatkowski, DJ [1 ]
Stossel, TP [1 ]
FuruUchi, K [1 ]
Kuzumaki, N [1 ]
机构
[1] HARVARD UNIV,BRIGHAM & WOMENS HOSP,SCH MED,DIV EXPT MED,BOSTON,MA 02115
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1997年 / 248卷 / 03期
关键词
gelsolin; cofilin; F-actin; severing inhibitor;
D O I
10.1111/j.1432-1033.1997.00834.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Gelsolin is a calcium-activated actin-binding protein with six subdomains. The N-terminal (G1) domain is essential for actin-filament-severing activity while other domains within G2-3 position the protein on the filament side allowing G1 to sever. In order to generate reagents capable of competitively inhibiting endogenous gelsolin and, potentially, other actin filament regulatory protein, we expressed several truncates of gelsolin in Escherichia coli, and analyzed how they affected the in vitro activity of two different actin-binding proteins, gelsolin and cofilin. A Ca2+-sensitive truncate containing G2-6 inhibited the F-actin-depolymerizing activities of both gelsolin and cofilin. while a G2-3 truncate was less effective. Using two independent assays, our results support the idea that gelsolin truncates inhibit actin filament severing and do not markedly affect actin subunit dissociation kinetics, Cosedimentation assays in the presence of calcium demonstrate that the G2-6 truncate binds to F-actin more strongly than the G2-3 truncate consistent with a protection mechanism by conformational change of F-actin and/or competitive binding to actin filaments which depends upon the presence of actin filament binding domains.
引用
收藏
页码:834 / 839
页数:6
相关论文
共 37 条
[1]  
ALLEN PG, 1994, J BIOL CHEM, V269, P32916
[2]  
Allen PG, 1996, J BIOL CHEM, V271, P4665
[3]   Dependence of fibroblast migration on actin severing activity of gelsolin [J].
Arora, PD ;
McCulloch, CAG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (34) :20516-20523
[4]   GELSOLIN HAS 3 ACTIN-BINDING SITES [J].
BRYAN, J .
JOURNAL OF CELL BIOLOGY, 1988, 106 (05) :1553-1562
[5]   Actin depolymerizing factor (ADF/cofilin) enhances the rate of filament turnover: Implication in actin-based motility [J].
Carlier, MF ;
Laurent, V ;
Santolini, J ;
Melki, R ;
Didry, D ;
Xia, GX ;
Hong, Y ;
Chua, NH ;
Pantaloni, D .
JOURNAL OF CELL BIOLOGY, 1997, 136 (06) :1307-1322
[6]   THE ACTIN FILAMENT SEVERING DOMAIN OF PLASMA GELSOLIN [J].
CHAPONNIER, C ;
JANMEY, PA ;
YIN, HL .
JOURNAL OF CELL BIOLOGY, 1986, 103 (04) :1473-1481
[7]   THE ROLE OF ACTIN POLYMERIZATION IN CELL MOTILITY [J].
COOPER, JA .
ANNUAL REVIEW OF PHYSIOLOGY, 1991, 53 :585-605
[8]   ENHANCED MOTILITY IN NIH-3T3 FIBROBLASTS THAT OVEREXPRESS GELSOLIN [J].
CUNNINGHAM, CC ;
STOSSEL, TP ;
KWIATKOWSKI, DJ .
SCIENCE, 1991, 251 (4998) :1233-1236
[9]   ISOLATION AND SOME STRUCTURAL AND FUNCTIONAL-PROPERTIES OF MACROPHAGE TROPOMYOSIN [J].
FATTOUM, A ;
HARTWIG, JH ;
STOSSEL, TP .
BIOCHEMISTRY, 1983, 22 (05) :1187-1193
[10]   FUNCTIONS OF [HIS321]GELSOLIN ISOLATED FROM A FLAT REVERTANT OF RAS-TRANSFORMED CELLS [J].
FUJITA, H ;
LAHAM, LE ;
JANMEY, PA ;
KWIATKOWSKI, DJ ;
STOSSEL, TP ;
BANNO, Y ;
NOZAWA, Y ;
MULLAUER, L ;
ISHIZAKI, A ;
KUZUMAKI, N .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1995, 229 (03) :615-620