A novel disulfide-rich protein motif from avian eggshell membranes.
A novel disulfide-rich protein motif from avian eggshell membranes.
复制标题
来自禽类蛋壳膜的新型富含二硫键的蛋白质基序。
DOI：
10.1371/journal.pone.0018187
复制
发表时间：
2011-03-30
期刊：
影响因子：
3.7
通讯作者：
Thorpe C
中科院分区：
文献类型：
--
作者：
Kodali VK;Gannon SA;Paramasivam S;Raje S;Polenova T;Thorpe C
在鸡蛋的壳下是两层相对的富含二硫化物的蛋白质膜。它们在禽类输卵管中使用由蛋白质形成的纤维制造，所述蛋白质通过不可逆的赖氨酸衍生的交联广泛偶联。这些蛋壳膜（ESM）的难处理性减缓了它们的表征，并且它们的蛋白质组成仍然不确定。在这项工作中，ESM的还原性烷基化，然后通过蛋白水解消化导致鉴定富含半胱氨酸的ESM蛋白（缩写为CREMP），其类似于来自细胞黏菌的孢子外壳蛋白SP 75。CREMP部分序列中的半胱氨酸重复序列的分析揭示了显着的重复模式运行。模块a含有C-X4-C-X5-C-X8-C-X6模式（其中X代表插入的非半胱氨酸残基）。这些半胱氨酸间氨基酸残基也是惊人地保守的。进化相关模块B具有与a相同的半胱氨酸间隔，但在其C-末端具有11个氨基酸残基。鸡基因组DNA片段中CREMP序列的不同延伸显示不同的重复模式：例如，所有a模块; a-B模块的交替;或a-B-B排列。在斑胸草雀（Taeniopygia guttata）和卵生绿色变色蜥蜴（Anolis carolinensis）的重叠群中发现了类似的CREMP蛋白。在所有这些情况下，高度保守的模块化重复序列的长运行显然导致全长DNA序列的组装困难。因此，CREMP蛋白的数量和氨基酸长度目前尚不清楚。来自鸡输卵管EST文库的118个氨基酸片段（代表a-b-a-b模式）在大肠杆菌中表达，是高度折叠的、高度各向异性的蛋白质，在2D溶液NMR谱中具有大的化学位移分散。该蛋白质片段的8个二硫键还原后结构完全丧失。最后，固态NMR光谱表明，在完整的ESM纤维令人惊讶的程度的秩序。
Under the shell of a chicken egg are two opposed proteinaceous disulfide-rich membranes. They are fabricated in the avian oviduct using fibers formed from proteins that are extensively coupled by irreversible lysine-derived crosslinks. The intractability of these eggshell membranes (ESM) has slowed their characterization and their protein composition remains uncertain. In this work, reductive alkylation of ESM followed by proteolytic digestion led to the identification of a cysteine rich ESM protein (abbreviated CREMP) that was similar to spore coat protein SP75 from cellular slime molds. Analysis of the cysteine repeats in partial sequences of CREMP reveals runs of remarkably repetitive patterns. Module a contains a C-X4-C-X5-C-X8-C-X6 pattern (where X represents intervening non-cysteine residues). These inter-cysteine amino acid residues are also strikingly conserved. The evolutionarily-related module b has the same cysteine spacing as a, but has 11 amino acid residues at its C-terminus. Different stretches of CREMP sequences in chicken genomic DNA fragments show diverse repeat patterns: e.g. all a modules; an alternation of a-b modules; or an a-b-b arrangement. Comparable CREMP proteins are found in contigs of the zebra finch (Taeniopygia guttata) and in the oviparous green anole lizard (Anolis carolinensis). In all these cases the long runs of highly conserved modular repeats have evidently led to difficulties in the assembly of full length DNA sequences. Hence the number, and the amino acid lengths, of CREMP proteins are currently unknown. A 118 amino acid fragment (representing an a-b-a-b pattern) from a chicken oviduct EST library expressed in Escherichia coli is a well folded, highly anisotropic, protein with a large chemical shift dispersion in 2D solution NMR spectra. Structure is completely lost on reduction of the 8 disulfide bonds of this protein fragment. Finally, solid state NMR spectra suggest a surprising degree of order in intact ESM fibers.
登录
查看更多内容
影响因子：
2.7
作者：
Gregg, KY;Cox, EC
通讯作者：
Cox, EC
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
--
作者：
HOFFER, AP
通讯作者：
HOFFER, AP
影响因子：
4.8
作者：
Hoober, KL;Sheasley, SL;Thorpe, C
通讯作者：
Thorpe, C
影响因子：
2.7
作者：
DELAGLIO, F;GRZESIEK, S;BAX, A
通讯作者：
BAX, A