title: A novel missense mutation of SLC7A9 frequent in Japanese cystinuria cases affecting the C-terminus of the transporter.
abstract: Cystinuria is caused by the inherited defect of apical membrane transport systems for cystine and dibasic amino acids in renal proximal tubules. Mutations in either SLC7A9 or SLC3A1 gene result in cystinuria. The mutations of SLC7A9 gene have been identified mainly from Italian, Libyan Jewish, North American, and Spanish patients. In the present study, we have analyzed cystinuria cases from oriental population (mostly Japanese). Mutation analyses of SLC7A9 and SLC3A1 genes were performed on 41 cystinuria patients. The uptake of 14C-labeled cystine in COS-7 cells was measured to determine the functional properties of mutants. The protein expression and localization were examined by Western blot and confocal laser-scanning microscopy. Among 41 patients analyzed, 35 were found to possess mutations in SLC7A9. The most frequent one was a novel missense mutation P482L that affects a residue near the C-terminus end of the protein and causes severe loss of function. In MDCK II and HEK293 cells, we found that P482L protein was expressed and sorted to the plasma membrane as well as wild type. The alteration of Pro482 with amino acids with bulky side chains reduced the transport function of b(0,+)AT/BAT1. Interestingly, the mutations of SLC7A9 for Japanese cystinuria patients are different from those reported for European and American population. The results of the present study contribute toward understanding the distribution and frequency of cystinuria-related mutations of SLC7A9.
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Shinkawa, Mitaka, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Shigeta
authors.forename: Y
authors.initials: Y
authors.lastname: Kanai
authors.forename: A
authors.initials: A
authors.lastname: Chairoungdua
authors.forename: N
authors.initials: N
authors.lastname: Ahmed
authors.forename: S
authors.initials: S
authors.lastname: Sakamoto
authors.forename: H
authors.initials: H
authors.lastname: Matsuo
authors.forename: D K
authors.initials: DK
authors.lastname: Kim
authors.forename: M
authors.initials: M
authors.lastname: Fujimura
authors.forename: N
authors.initials: N
authors.lastname: Anzai
authors.forename: K
authors.initials: K
authors.lastname: Mizoguchi
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: K
authors.initials: K
authors.lastname: Akakura
authors.forename: T
authors.initials: T
authors.lastname: Ichikawa
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: H
authors.initials: H
authors.lastname: Endou
mesh_terms: D019943:Amino Acid Substitution; D026923:Amino Acid Transport Systems, Basic / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003555:Cystinuria / Q000235:genetics* / Q000473:pathology; D017931:DNA Primers; D005091:Exons; D006580:Genetic Carrier Screening; D006720:Homozygote; D006801:Humans; D007438:Introns; D007564:Japan; D018613:Microscopy, Confocal; D008958:Models, Molecular; D020125:Mutation, Missense; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection
chemical_list: D026923:Amino Acid Transport Systems, Basic; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; C499489:SLC7A9 protein, human