Sparrow L G, Robinson C P, Caine J, McMahon D T, Strike P M
C.S.I.R.O. Division of Wool Technology, Parkville, Vic., Australia.
Biochem J. 1992 Feb 15;282 ( Pt 1)(Pt 1):291-7. doi: 10.1042/bj2820291.
Component 5 is one of the four type II intermediate-filament proteins found in the hard keratin wool. It was isolated as the S-carboxymethyl derivative from Merino wool and its amino acid sequence was determined by manual and automatic sequencing of peptides produced by chemical and enzymic cleavage. Component 5 is an N-terminally blocked molecule of 503 residues and Mr (not including the blocking group) of 56,600. The blocking group has not been identified. The amino acid sequence of component 5 shows 77% sequence identity with that of component 7c, another type II wool intermediate-filament protein [Sparrow, Robinson, McMahon & Rubira (1989) Biochem. J. 261, 1015-1022]. The sequence similarity extends from the N-termini of the two molecules to residue 459 (component 5 sequence); however, there is no recognizable sequence similarity in the remaining C-terminal 43 amino acid residues. Details of procedures used in determining the sequence of component 5 have been deposited as a Supplementary Publication SUP 50168 (80 pages) at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire, LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1992) 281, 5. The information comprises: (1) details of chemical and enzymic methods used for cleavage of component 5, peptide CN1, the peptide mixture CN2/3 and various other peptides, (2) details of the procedures used for the fractionation and purification of peptides from (1), including Figures showing the elution profiles from the chromatographic steps used, and (3) details of the method used to determine the C-terminal sequence of component 5.
组分5是在硬角蛋白羊毛中发现的四种II型中间丝蛋白之一。它是从美利奴羊毛中分离出来的S-羧甲基衍生物,其氨基酸序列通过对化学和酶切产生的肽段进行手动和自动测序来确定。组分5是一个N端封闭的分子,有503个残基,Mr(不包括封闭基团)为56,600。封闭基团尚未确定。组分5的氨基酸序列与另一种II型羊毛中间丝蛋白组分7c的氨基酸序列有77%的序列同一性[斯帕罗、罗宾逊、麦克马洪和鲁比拉(1989年)《生物化学杂志》261卷,1015 - 1022页]。序列相似性从两个分子的N端延伸到第459位残基(组分5序列);然而,在其余的C端43个氨基酸残基中没有可识别的序列相似性。确定组分5序列所使用程序的详细信息已作为补充出版物SUP 50168(80页)存放在英国西约克郡韦瑟比波士顿温泉市的大英图书馆文献供应中心,邮编LS23 7BQ,可按《生物化学杂志》(1992年)281卷第5期所示条件从该处获取副本。这些信息包括:(1)用于切割组分5、肽CN1、肽混合物CN2/3和各种其他肽的化学和酶法的详细信息,(2)用于从(1)中分离和纯化肽的程序的详细信息,包括显示所用色谱步骤洗脱图谱的图,以及(3)用于确定组分5 C端序列的方法的详细信息。