Mutational analysis of type IV collagen alpha5 chain, with respect to heterotrimer formation.
Alport syndrome (AS) is caused by mutations in type IV collagen alpha3, alpha4, and alpha5 chains. The three chains form a heterotrimer. In this study, we introduced 12 kinds of missense and three kinds of nonsense mutations, corresponding to AS mutations, into the NC1 domain of alpha5(IV) and characterized the mutant chains. Nine alpha5(IV) chains with amino acid substitutions and all three truncated alpha5(IV) chains did not form a heterotrimer and were not secreted from cells. Three alpha5(IV) chains with amino acid substitutions did, however, form heterotrimers in cells, but these were not secreted from cells. These findings indicate that a defect in heterotrimer formation is the main molecular mechanism underlying the pathogenesis of AS caused by mutation in the NC1 domain. We also showed that even a single amino acid deletion in the carboxyl-terminal region markedly affected the heterotrimerization, indicating that the carboxyl-terminal end is indispensable for heterotrimer formation.
Kobayashi
Takehiro
T
Division of Pediatrics, Department of Homeostatic Regulation and Development, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Niigata City 951-8510, Japan.
Kakihara
Toshio
T
Uchiyama
Makoto
M
D019943:Amino Acid Substitution; D001665:Binding Sites; D002460:Cell Line; D024141:Collagen Type IV / Q000737:chemistry* / Q000378:metabolism*; D019281:Dimerization; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding
C478159:COL4A5 protein, human; D024141:Collagen Type IV