Properties of GST-CALM expressed in E. coli.
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.
Kim
J A
JA
Department of Biochemistry, Medical College, Ewha Womans University, Seoul, Korea.
Kim
S R
SR
Jung
Y K
YK
Woo
S Y
SY
Seoh
J Y
JY
Hong
Y S
YS
Kim
H L
HL
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D000911:Antibodies, Monoclonal; D002154:Calpain / Q000737:chemistry; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000737:chemistry; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005982:Glutathione Transferase / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D034041:Monomeric Clathrin Assembly Proteins; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018909:src Homology Domains
D033942:Adaptor Proteins, Vesicular Transport; D000911:Antibodies, Monoclonal; D034041:Monomeric Clathrin Assembly Proteins; D009419:Nerve Tissue Proteins; C439003:PICALM protein, human; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C439023:clathrin assembly protein AP180; D005982:Glutathione Transferase; C505407:CASP3 protein, human; C505530:CASP8 protein, human; C506433:CASP9 protein, human; D002154:Calpain; C505409:Casp3 protein, mouse; C505526:Casp8 protein, mouse; C506434:Casp9 protein, mouse; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases