Properties of GST-CALM expressed in E. coli.
- Author:
Jeong Ah KIM
1
;
Seong Ryul KIM
;
Yong Keun JUNG
;
So Youn WOO
;
Ju Young SEOH
;
Young Sook HONG
;
Hyung Lae KIM
Author Information
1. Department of Biochemistry, Medical College, Ewha Womans University, Seoul, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
expression;
clathrin-coated vesicle;
CALM;
AP180;
regulation;
SH3 domain
- MeSH:
Animal;
Antibodies, Monoclonal;
Calpain/chemistry;
Caspases/chemistry;
Clathrin-Coated Vesicles/metabolism*;
Electrophoresis, Polyacrylamide Gel;
Escherichia coli/metabolism;
Escherichia coli/genetics;
Female;
Glutathione Transferase/genetics*;
Mice;
Mice, Inbred BALB C;
Nerve Tissue Proteins/metabolism;
Nerve Tissue Proteins/metabolism;
Nerve Tissue Proteins/chemistry*;
Phosphoproteins/metabolism;
Phosphoproteins/genetics;
Phosphoproteins/chemistry*;
Protein Binding;
Rabbits;
Recombinant Fusion Proteins/metabolism;
Recombinant Fusion Proteins/genetics;
Recombinant Fusion Proteins/chemistry*;
src Homology Domains
- From:Experimental & Molecular Medicine
2000;32(2):93-99
- CountryRepublic of Korea
- Language:English
-
Abstract:
Clathrin-coated vesicles (CCVs) are involved in protein and lipid trafficking between intracellular compartments in eukaryotic cells. CCVs are composed of clathrin and assembly proteins. The clathrin assembly protein lymphoid myeloid leukemia (CALM) gene, encodes a homologoue of the neuronal clathrin assembly protein AP180. In this study, we characterized the properties of the CALM expressed in E. coli. The molecular weight of bacterially expressed GST-CALM fusion protein was approximately 105 kD on SDS-PAGE. The CALM protein could promote clathrin triskelia into clathrin cages and could bind the preformed clathrin cage. However, 33 kD N-terminal domain of CALM could not bind pre-assembled clathrin cages, but assemble clathrin triskelia into clathrin cages. The CALM protein was bound to SH3 domain through N-terminal domain1, in vitro. The CALM protein is proteolyzed by caspase 3, caspase 8 and calpain through C-terminal domain.
