Division of Child and Adolescent Health, Department of Medical Genetics, University Hospital of North-Norway, Tromsø, Norway.
PLoS One. 2011;6(8):e22968. doi: 10.1371/journal.pone.0022968. Epub 2011 Aug 23.
Limb-Girdle Muscular Dystrophy type 2I (LGMD2I) is an inheritable autosomal, recessive disorder caused by mutations in the FuKutin-Related Protein (FKRP) gene (FKRP) located on chromosome 19 (19q13.3). Mutations in FKRP are also associated with Congenital Muscular Dystrophy (MDC1C), Walker-Warburg Syndrome (WWS) and Muscle Eye Brain disease (MEB). These four disorders share in common an incomplete/aberrant O-glycosylation of the membrane/extracellular matrix (ECM) protein α-dystroglycan. However, further knowledge on the FKRP structure and biological function is lacking, and its intracellular location is controversial. Based on immunogold electron microscopy of human skeletal muscle sections we demonstrate that FKRP co-localises with the middle-to-trans-Golgi marker MG160, between the myofibrils in human rectus femoris muscle fibres. Chemical cross-linking experiments followed by pairwise yeast 2-hybrid experiments, and co-immune precipitation, demonstrate that FKRP can exist as homodimers as well as in large multimeric protein complexes when expressed in cell culture. The FKRP homodimer is kept together by a disulfide bridge provided by the most N-terminal cysteine, Cys6. FKRP contains N-glycan of high mannose and/or hybrid type; however, FKRP N-glycosylation is not required for FKRP homodimer or multimer formation. We propose a model for FKRP which is consistent with that of a Golgi resident type II transmembrane protein.
肢带型肌营养不良 2I 型(LGMD2I)是一种由 FuKutin 相关蛋白(FKRP)基因突变引起的常染色体隐性遗传性疾病,该基因位于 19 号染色体(19q13.3)上。FKRP 基因突变也与先天性肌营养不良症(MDC1C)、Walker-Warburg 综合征(WWS)和肌肉眼脑疾病(MEB)有关。这四种疾病的共同特征是膜/细胞外基质(ECM)蛋白α- dystroglycan 的不完全/异常 O-糖基化。然而,对于 FKRP 的结构和生物学功能的进一步了解还很缺乏,其细胞内定位也存在争议。基于对人类骨骼肌切片的免疫金电子显微镜研究,我们证明 FKRP 与中间到转高尔基标记物 MG160 共定位,位于人类股直肌纤维的肌原纤维之间。化学交联实验随后进行酵母 2 杂交实验和共免疫沉淀实验,证明 FKRP 可以作为同源二聚体存在,也可以在细胞培养中形成大的多聚体蛋白复合物。FKRP 同源二聚体由最 N 端半胱氨酸 Cys6 提供的二硫键保持在一起。FKRP 含有高甘露糖和/或杂合型的 N-聚糖;然而,FKRP 的 N-糖基化对于 FKRP 同源二聚体或多聚体的形成不是必需的。我们提出了一个与高尔基驻留型 II 型跨膜蛋白一致的 FKRP 模型。