Robbins P W, Trimble R B, Wirth D F, Hering C, Maley F, Maley G F, Das R, Gibson B W, Royal N, Biemann K
J Biol Chem. 1984 Jun 25;259(12):7577-83.
We report the DNA and primary amino acid sequences of the Streptomyces plicatus enzyme endo-beta-N-acetylglucosaminidase H. Peptide sequence information was derived from enzyme isolated from Streptomyces culture medium using a combination of mass spectrometric methods and conventional techniques, including Edman degradation and carboxypeptidase Y digestion. The DNA sequence was determined by analysis of the Endo-beta-N-acetylglucosaminidase H gene cloned into the Escherichia coli plasmid pBR322 (Robbins, P. W., Wirth , D. F., and Hering , C. (1981) J. Biol. Chem. 256, 10640-10644). The enzyme from Streptomyces medium is 271 (or 269) amino acids in length and has a ragged NH2-terminal sequence beginning primarily with Ala-Pro-Val or Ala-Pro-Ala-Pro-Val. DNA resection experiments as well as the DNA sequence itself suggest that a proenzyme or, more probably, " prepro " enzyme may be the primary product of translation. The long 42 (or 44) residue leader sequence of the preproenzyme shows striking similarities to leader sequences found on proteins secreted by Bacillus species. The leader sequence is partially removed by E. coli and, as reported previously, endo-beta-N-acetylglucosaminidase H made in E. coli appears in both the periplasmic space and in the cell.
我们报道了褶皱链霉菌内切-β-N-乙酰氨基葡萄糖苷酶H的DNA和一级氨基酸序列。肽序列信息来自于使用质谱方法和传统技术（包括埃德曼降解和羧肽酶Y消化）从链霉菌培养基中分离得到的酶。通过分析克隆到大肠杆菌质粒pBR322中的内切-β-N-乙酰氨基葡萄糖苷酶H基因来确定DNA序列（罗宾斯，P.W.，沃思，D.F.，和赫林，C.（1981年）《生物化学杂志》256，10640 - 10644）。来自链霉菌培养基的酶长度为271（或269）个氨基酸，其NH2末端序列参差不齐，主要以丙氨酸-脯氨酸-缬氨酸或丙氨酸-脯氨酸-丙氨酸-脯氨酸-缬氨酸开头。DNA切除实验以及DNA序列本身表明，酶原或更有可能是“前酶原”可能是翻译的主要产物。前酶原的长42（或44）个残基的前导序列与芽孢杆菌属分泌的蛋白质上的前导序列有显著相似性。前导序列被大肠杆菌部分切除，并且如先前报道的那样，在大肠杆菌中产生的内切-β-N-乙酰氨基葡萄糖苷酶H出现在周质空间和细胞中。
