Amino acid sequence of human lens beta B2-crystallin.
Amino acid sequence of human lens beta B2-crystallin.
复制标题
人晶状体 β B2-晶状体蛋白的氨基酸序列。
DOI：
10.1002/pro.5560020217
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Smith,DL
中科院分区：
文献类型：
--
作者：
Miesbauer,LR;Smith,JB;Smith,DL
晶状体细胞核中的蛋白质和生物体一样古老，这使得晶状体成为研究生长和衰老的有用模型。此外，晶状体蛋白的修饰和交联会引起人们的兴趣，因为它们被认为在白内障的形成中起着重要作用。哺乳动物的晶状体包含三个结构蛋白家族，a-、P-和y-晶体蛋白。在这些化合物中，0-晶状体蛋白的异质性最强，特性也最差。只有两个人0-晶体蛋白的初级序列，PA3/A1和PB3(部分序列)，已经从他们的基因组DNA序列中被确定(Hogg等人，1986年；Aart等人，1989a)。虽然牛、大鼠和小鼠已知0-晶体蛋白主要成分Pb2的氨基酸序列，但只有一小部分人类基因序列已被报道(Driessen等人，1981年；Inana等人，1982年；Aart等人，198913；Chambers&Russell，1991年)。通过检测人类白内障晶状体中由钙依赖的谷氨酰胺转氨酶介导的交联物形成的蛋白质聚合物，已经证明了@B2在白内障发生中的可能作用(Lorand等人，1981)。对兔晶状体的进一步研究表明，这种交联物可与β-晶状体蛋白产生(不与α-或γ-晶状体蛋白)，并可被牛PBP抗血清识别(Vela co&Lorand，1987)。为了确定这些交联链和其他可能存在于白内障晶状体中的交联链的性质，PB2的氨基酸序列是必要的。我们利用与牛的序列同源性，对人PB2进行了胰酶和内蛋白酶Glu-C多肽的质谱分析。正常人晶状体的水溶部分通过Sephadex G-200凝胶过滤层析分离得到α-、6-和y-晶体蛋白。@-晶体蛋白部分通过反相高效液相色谱分离，在C4色谱柱上使用水/乙腈(0.1070.3%三氟乙酸)
The proteins in the lens nucleus are as old as the organism, making the lens a useful model in which to study growth and aging. Furthermore, modification and crosslinking of the lens proteins are of interest because they are thought to be important in cataract formation. The mammalian lens contains three families of structural proteins, the a-, P-, and y-crystallins. Of these, the 0-crystallins are the most heterogeneous and the least well characterized. The primary sequences of only two human 0-crystallins, PA3/Al and PB3 (partial sequence), have been determined from their genomic DNA sequences (Hogg et al., 1986; Aarts et al., 1989a). Although the amino acid sequence of the principal component of the 0-crystallins, PB2, formerly called PBp, is known for bovine, rat and mouse, only a small section of the human gene sequence has been reported (Driessen et al., 1981; Inana et al., 1982; Aarts et al., 198913; Chambers & Russell, 1991). A possible role of@ B2 in cataractogenesis has been demonstrated by detection of a protein polymer in human cataractous lenses, formed by a Ca’+-dependent transglutaminase mediated cross-link (Lorand et al., 1981). Further studies with rabbit lenses showed that this crosslink could be produced with P-crystallin (not with a-or y-crystallin) and was recognized by antiserum to bovine PBp (Velasco & Lorand, 1987). To identify the nature of those cross-links and others that may be present in cataractous lenses, the amino acid sequence of PB2 is necessary. We have sequenced human PB2 by mass spectrometric analysis of trypsin and endoproteinase Glu-C-generated peptides using homology to the bovine sequence as a reference.The water-soluble portion of a normal human lens was fractionated into a-, 6-, and y-crystallins by Sephadex G-200 gel filtration chromatography.@ B2 was isolated from the@-crystallin fraction by reversed-phase high performance liquid chromatography (HPLC) on a C4 column using a water/acetonitrile (0.1070 trifluoroacetic acid