Possible involvement of clathrin in neuritogenesis induced by a protease inhibitor (benzyloxycarbonyl-Leu-Leu-Leu-aldehyde) in PC12 cells.
Our previous reports showed that benzyloxycarbonyl (Z)-Leu-Leu-Leu-al (ZLLLal) induces neurite outgrowth in PC12 cells, and that 33-, 35-, and 180-kDa proteins from PC12 cells elute specifically from a Leu-Leu-Leu-al (LLLal)-coupled affinity column. Several lines of evidence suggest that the 33-, 35-, and 180-kDa proteins are components of clathrin, well-known for its role in endocytosis. Separation of clathrin into its heavy and light chains showed that the clathrin heavy chains have the ability to bind to a LLLal affinity column directly. Furthermore, ZLLLal enhances the rate of polymerization of clathrin triskelion to the coat structure. ZLLL-COOH does not cause neurite outgrowth in PC12 cells, and has no effect on the rate of clathrin polymerization. On immunocytochemical analysis of PC12 cells with an anti-clathrin heavy chain antibody, enhanced staining of the clathrin heavy chain was observed concomitantly with neurite outgrowth initiated by ZLLLal, but not by NGF. This study provides new insights into both the role of the clathrin molecule and the regulatory mechanism of neurite outgrowth.
Saito
Y
Y
Department of Enzyme Biochemistry, Tokyo Metropolitan Institute of Gerontology.
Tsubuki
S
S
Ito
H
H
Ohmi-Imajo
S
S
Kawashima
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002846:Chromatography, Affinity; D002966:Clathrin / Q000302:isolation & purification / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D010450:Endopeptidases / Q000187:drug effects / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology*; D008099:Liver / Q000737:chemistry; D046911:Macromolecular Substances; D008954:Models, Biological; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000502:physiology; D016501:Neurites / Q000187:drug effects / Q000502:physiology*; D016716:PC12 Cells; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
D002966:Clathrin; D007976:Leupeptins; D046911:Macromolecular Substances; D009414:Nerve Growth Factors; D011480:Protease Inhibitors; D010450:Endopeptidases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde