Properties of GST-CALM expressed in E. coli.

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.
Authors	J A Kim, S R Kim, Y K Jung, S Y Woo, J Y Seoh, Y S Hong, H L Kim
Journal	Experimental & molecular medicine (Exp Mol Med) Vol. 32 Issue 2 Pg. 93-9 (Jun 30 2000) ISSN: 1226-3613 [Print] United States
PMID	10926122 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Adaptor Proteins, Vesicular Transport Antibodies, Monoclonal Monomeric Clathrin Assembly Proteins Nerve Tissue Proteins PICALM protein, human Phosphoproteins Recombinant Fusion Proteins clathrin assembly protein AP180 Glutathione Transferase CASP3 protein, human CASP8 protein, human CASP9 protein, human Calpain Casp3 protein, mouse Casp8 protein, mouse Casp9 protein, mouse Caspase 3 Caspase 8 Caspase 9 Caspases
Topics	Adaptor Proteins, Vesicular Transport Animals Antibodies, Monoclonal Calpain (chemistry) Caspase 3 Caspase 8 Caspase 9 Caspases (chemistry) Clathrin-Coated Vesicles (metabolism) Electrophoresis, Polyacrylamide Gel Escherichia coli (genetics, metabolism) Female Glutathione Transferase (genetics) Mice Mice, Inbred BALB C Monomeric Clathrin Assembly Proteins Nerve Tissue Proteins (chemistry, genetics, metabolism) Phosphoproteins (chemistry, genetics, metabolism) Protein Binding Rabbits Recombinant Fusion Proteins (chemistry, genetics, metabolism) src Homology Domains

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