The consequences of deglycosylation of recombinant intra-melanosomal domain of human tyrosinase

被引：8

作者：

Dolinska, Monika B. ^{[1
]}

Sergeev, Yuri V. ^{[1
]}

机构：

[1] NEI, Ophthalm Genet & Visual Funct Branch, NIH, 10 Ctr Dr,31 Ctr Dr MSC 2510, Bethesda, MD 20892 USA

来源：

BIOLOGICAL CHEMISTRY | 2018年 / 399卷 / 01期

关键词：

albinism; genetic mutations; N-glycosylation; post-translational modification; protein activity; protein purification; ER-ASSOCIATED DEGRADATION; OCULOCUTANEOUS ALBINISM; MELANOMA-CELLS; N-GLYCOSYLATION; MUTATIONS; SITE;

D O I：

10.1515/hsz-2017-0178

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Tyrosinase, a melanosomal glycoenzyme, catalyzes initial steps of the melanin biosynthesis. While glycosylation was previously studied in vivo, we present three recombinant mutant variants of human tyrosinase, which were obtained using multiple site-directed mutagenesis, expressed in insect larvae, purified and characterized biochemically. The mutagenesis demonstrated the reduced protein expression and enzymatic activity due to possible loss of protein stability and protein degradation. However, the complete deglycosylation of asparagine residues in vitro, including the residue in position 371, interrupts tyrosinase function, which is consistent with a melanin loss in oculocutaneous albinism type 1 (OCA1) patients.

引用

页码：73 / 77

页数：5

共 16 条

[1] Mutations at critical N-glycosylation sites reduce tyrosinase activity by altering folding and quality control [J].