CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF A GENE ENCODING AN ADHESIN SUBUNIT PROTEIN OF HELICOBACTER-PYLORI
CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF A GENE ENCODING AN ADHESIN SUBUNIT PROTEIN OF HELICOBACTER-PYLORI
复制标题
DOI：
10.1128/jb.175.3.674-683.1993
复制
发表时间：
1993-02-01
影响因子：
3.2
通讯作者：
LEE, CH
中科院分区：
文献类型：
--
作者：
EVANS, DG;KARJALAINEN, TK;LEE, CH
克隆了幽门螺杆菌N-乙酰神经氨酸乳糖结合纤维血凝素（NLBH）受体结合亚单位hpaA基因，并进行了序列测定。由hpaA表达的蛋白，命名为HpaA，根据其胎球蛋白结合活性及其与从H.幽门。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析和蛋白质印迹（免疫印迹）表明，克隆的粘附素与H.幽门。此外，HpaA含有一个短的氨基酸序列（KRTIQK），这些氨基酸序列与构成大肠杆菌SfaS、K99和CFA/I的唾液酸结合基序的氨基酸序列相同或功能相似。亲和纯化的抗体对包含该序列的12个残基的合成肽具有特异性，可阻断H. pylori感染，免疫金电镜显示与未染色的H. pylori细胞，这与先前关于NLBH的位置和形态的观察一致。
Gene hpaA, which codes for the receptor-binding subunit of the N-acetylneuraminyllactose-binding fibrillar hemagglutinin (NLBH) of Helicobacter pylori, was cloned and sequenced. The protein expressed by hpaA, designated HpaA, was identified as the adhesin subunit on the basis of its fetuin-binding activity and its reactivity with a polyclonal, monospecific rabbit serum prepared against NLBH purified from H. pylori. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Western blots (immunoblots) showed that the cloned adhesin has the same molecular weight (20,000) as that found on H. pylori. Also, HpaA contains a short sequence of amino acids (KRTIQK) which are all either identical or functionally similar to those which compose the sialic acid-binding motif of Escherichia coli SfaS, K99, and CFA/I. Affinity-purified antibody specific for a 12-residue synthetic peptide that included this sequence blocked the hemagglutinating activity of H. pylori and was shown by immuno-gold electron microscopy to react with almost transparent material on unstained H. pylori cells, which is consistent with previous observations concerning the location and morphology of the NLBH.