Purification of clathrin assembly protein from rat liver.
Recently, the gene encoding clathrin assembly protein of lymphoid myeloid leukemia (CALM), which is homologous to the AP180, was cloned from rat brain, and its expression differential to AP180 was reported (Kim and Lee, 1999). This gene product promotes the polymerization of clathrin into clathrin cage and found to be a regulator in membrane trafficking between intracellular compartments in eukaryotic cells (Kim et al., 2000). In this study, we have purified the CALM protein from clathrin-coated vesicles of rat liver using the monoclonal antibody against the recombinant N-terminal region of the CALM. The coated proteins extracted from the coated vesicle fraction was further purified by multi-step procedures involving gel-filtration and ion-exchange chromatography and SDS-PAGE. The purified protein with an apparent molecular weight of 100 kD promoted the assembly of clathrin triskelia into clathrin cage. In this respect the CALM protein bears a functional resemblance to the AP180 that has been previously described.
Kim
H L
HL
Department of Biochemistry, Medical College, Ewha Womans University, Seoul, Korea. hyung@mm.ewha.ac.kr
Kim
J A
JA
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002966:Clathrin / Q000378:metabolism*; D022163:Clathrin-Coated Vesicles / Q000737:chemistry*; D008099:Liver / Q000737:chemistry*; D034041:Monomeric Clathrin Assembly Proteins; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D010750:Phosphoproteins / Q000302:isolation & purification*; D051381:Rats
D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D034041:Monomeric Clathrin Assembly Proteins; D009419:Nerve Tissue Proteins; C439003:PICALM protein, human; D010750:Phosphoproteins; C439023:clathrin assembly protein AP180