Wang J, Karabinos A, Schünemann J, Riemer D, Weber K
Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, Germany.
Eur J Cell Biol. 2000 Jul;79(7):478-87. doi: 10.1078/0171-9335-00069.
Two novel cytoplasmic intermediate filament (IF) proteins (C and D) from the tunicate (urochordate) Styela are characterised as putative keratin orthologs. The coexpression of C and D in all epidermal cells and the obligatory heteropolymeric IF assembly of the recombinant proteins argue for keratin orthologs, but the sequences do not directly reveal which protein behaves as a keratin I or II ortholog. This problem is solved by the finding that keratin 8, a type II keratin from man or Xenopus, forms chimeric IF when mixed with Styela D. Mutant proteins of Styela D and keratin 8 with a single cysteine in equivalent positions show that these chimeric IF are, like vertebrate keratin filaments, based on the hetero coiled coil. We propose that Styela D retains, in spite of its strong sequence drift, important molecular features of type I keratins. By inference Styela C reflects a type II ortholog. We discuss that type I to III IF proteins are expressed along the chordate branch of metazoa.
来自被囊动物(尾索动物)柄海鞘的两种新型细胞质中间丝(IF)蛋白(C和D)被鉴定为假定的角蛋白直系同源物。C和D在所有表皮细胞中的共表达以及重组蛋白的强制性异聚IF组装支持角蛋白直系同源物的观点,但序列并未直接揭示哪种蛋白表现为角蛋白I或II的直系同源物。通过发现人或非洲爪蟾的II型角蛋白角蛋白8与柄海鞘D混合时形成嵌合中间丝,这个问题得到了解决。柄海鞘D和角蛋白8在等效位置具有单个半胱氨酸的突变蛋白表明,这些嵌合中间丝与脊椎动物角蛋白丝一样,基于异源卷曲螺旋。我们提出,尽管柄海鞘D的序列有很大漂移,但它保留了I型角蛋白的重要分子特征。由此推断,柄海鞘C反映了II型直系同源物。我们讨论了I至III型中间丝蛋白在后生动物的脊索动物分支中表达。