Disulfide-bonding between Drosophila laminin β and γ chains is essential for α chain to form αβγ trimer

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• 作者：Kumagai ; Chino ; Kadowaki ; Tatsuhiko ; Kitagawa ; Yasuo
• 关键词：Basement membrane ; Disulfide-bonding ; Drosophila ; Laminin ; CBB ; Coomassie brilliant blue ; ECL ; enhanced chemiluminescence ; EGF ; epidermal growth factor ; EHS ; Engelbreth-Holm-Swarm ; NEM ; N-ethylmaleimide ; PBS ; phosphate-buffered saline ; RIPA ; radio-immunop
• 刊名：FEBS Letters
• 出版年：1997
• 出版时间：July 21, 1997
• 年：1997
• 卷：412
• 期：1
• 页码：211-216
• 全文大小：1.16 M

文摘

Assembly of Drosophila laminin α, β and γ chains was analyzed by immunoprecipitation of the lysate from metabolically radiolabeled Kc 167 cells with chain-specific antibodies followed by two dimensional electrophoresis in which non-reducing and reducing SDS gel electrophoresis are combined. Precipitation of monomeric β (or γ) with anti-γ (or -β) antibody revealed that β and γ form stable dimer before they are disulfide-bonded to each other. In contrast, α associates with neither monomeric β, monomeric γ nor βγ dimer without disulfide-bonding but only with disulfide-bonded βγ dimer to form αβγ trimers. These results thus demonstrated that the interchain disulfide-boding between β and γ is essential for α to form αβγ trimer. We also found that the αβγ trimer can be secreted with α chain either disulfide-bonded or not bonded to the disulfide-bonded βγ dimer.