CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF THE PASTEURELLA-HAEMOLYTICA A1 GLYCOPROTEASE GENE
CLONING, NUCLEOTIDE-SEQUENCE, AND EXPRESSION OF THE PASTEURELLA-HAEMOLYTICA A1 GLYCOPROTEASE GENE
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DOI：
10.1128/jb.173.18.5597-5603.1991
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发表时间：
1991-09-01
影响因子：
3.2
通讯作者：
MELLORS, A
中科院分区：
文献类型：
--
作者：
ABDULLAH, KM;LO, RYC;MELLORS, A
溶血巴斯德氏菌血清型A1分泌一种糖蛋白酶，其对O-唾液酸糖蛋白如血型糖蛋白A具有特异性。已将编码糖蛋白酶的基因克隆到重组质粒pPH 1中，并测定了其核苷酸序列。该基因（命名为gcp）编码35.2 kDa的蛋白质，在携带pPH 1的大肠杆菌克隆中可以观察到该分子量的活性酶蛋白。在E.大肠杆菌maxicells证实了来自pPH 1的35-kDa蛋白的表达。异源表达的蛋白质的氨基末端序列对应于从核苷酸序列预测的序列。该糖蛋白酶是一种中性金属蛋白酶，并且该糖蛋白酶的预测氨基酸序列含有推定的锌结合位点。该基因与原核或真核来源的其他蛋白酶基因没有显着的同源性。然而，gcp和E.大肠杆菌基因orfX，其产物被认为在大分子生物合成的调节中起作用。
Pasteurella haemolytica serotype A1 secretes a glycoprotease which is specific for O-sialogycoproteins such as glycophorin A. The gene encoding the glycoprotease enzyme has been cloned in the recombinant plasmid pPH1, and its nucleotide sequence has been determined. The gene (designated gcp) codes for a protein of 35.2 kDa, and an active enzyme protein of this molecular mass can be observed in Escherichia coli clones carrying pPH1. In vivo labeling of plasmid-encoded proteins in E. coli maxicells demonstrated the expression of a 35-kDa protein from pPH1. The amino-terminal sequence of the heterologously expressed protein corresponds to that predicted from the nucleotide sequence. The glycoprotease is a neutral metalloprotease, and the predicted amino acid sequence of the glycoprotease contains a putative zinc-binding site. The gene shows no significant homology with the genes for other proteases of procaryotic or eucaryotic origin. However, there is substantial homology between gcp and an E. coli gene, orfX, whose product is believed to function in the regulation of macromolecule biosynthesis.