Chintamaneni C D, Halaban R, Kobayashi Y, Witkop C J, Kwon B S
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202-5120.
Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5272-6. doi: 10.1073/pnas.88.12.5272.
We have determined a molecular defect to be the likely basis for inactivity of the tyrosinase (EC 1.14.18.1) from a patient with tyrosinase-negative oculocutaneous albinism. A single base (thymine) was inserted in exon 5 of the tyrosinase gene following codon 471 in the putative transmembrane coding region. This insertion caused a shift in the reading frame of 19 amino acids at the 3' end and introduced a premature termination signal that would be expected to truncate the protein by 21 amino acids at the carboxyl terminus. The albino tyrosinase was not recognized by antibodies directed to the carboxyl terminus of tyrosinase. Furthermore, as shown by gel electrophoresis of the immunoprecipitated protein, the tyrosinase was approximately 3 kDa smaller than normal. Similar immunoprecipitation data were obtained when cloned normal and mutant tyrosinases were expressed in COS-1 cells.
我们已经确定,一名酪氨酸酶阴性眼皮肤白化病患者的酪氨酸酶(EC 1.14.18.1)无活性的可能原因是一种分子缺陷。在假定的跨膜编码区域中,第471密码子之后,酪氨酸酶基因的外显子5中插入了一个单碱基(胸腺嘧啶)。这种插入导致3'端阅读框发生19个氨基酸的移码,并引入了一个提前终止信号,预计该信号会使蛋白质在羧基末端截短21个氨基酸。针对酪氨酸酶羧基末端的抗体无法识别白化病患者的酪氨酸酶。此外,免疫沉淀蛋白的凝胶电泳显示,该酪氨酸酶比正常的小约3 kDa。当在COS-1细胞中表达克隆的正常和突变酪氨酸酶时,也获得了类似的免疫沉淀数据。