Exp Mol Med.  2000 Jun;32(2):93-99.

Properties of GST-CALM expressed in E. coli

• Kim JA
• Kim SR
• Jung YK
• Woo SY
• Seoh JY
• Hong YS
• Kim HL

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.

Keyword

expression; clathrin-coated vesicle; CALM; AP180; regulation; SH3 domain

MeSH Terms

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