COMPLETE AMINO-ACID SEQUENCE OF P453-PLASMID-MEDIATED PIT-2 BETA-LACTAMASE (SHV-1)
COMPLETE AMINO-ACID SEQUENCE OF P453-PLASMID-MEDIATED PIT-2 BETA-LACTAMASE (SHV-1)
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DOI：
10.1042/bj2510073
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发表时间：
1988-04-01
影响因子：
4.1
通讯作者：
LABIA, R
中科院分区：
文献类型：
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作者：
BARTHELEMY, M;PEDUZZI, J;LABIA, R
p453质粒介导的PIT-2 β-测定SHV-1酶活性。该蛋白质含有265个残基。用胰蛋白酶、金黄色葡萄球菌V_8蛋白酶、糜蛋白酶和Lys-C蛋白酶消化，然后用溴化氰裂解，用反相高效液相色谱法分离纯化肽。用异硫氰酸二甲氨基偶氮苯酯/异硫氰酸苯酯双偶联法手动测定每个肽的氨基酸序列。PIT-2 β-的一级结构将内酰胺酶与两种密切相关的酶，即TEM-1 β-内酰胺酶进行了比较。内酰胺酶和β-内酰胺酶肺炎克雷伯菌菌株LEN-1的内酰胺酶。PIT-2 β-内酰胺酶氨基酸序列保留较好，同源性分别为68%和88%。因此，PIT-2酶可以代表染色体编码的β-葡聚糖酶之间的进化步骤。内酰胺酶和质粒介导的TEM β-内酰胺酶。
The complete amino acid sequence of the p453-plasmid-mediated PIT-2 .beta.-lactamase (SHV-1) was determined. The protein contains 265 residues. Peptides resulting from digestions with trypsin, Staphylococcus aureus V8 proteinase, chymotrypsin and Lys-C proteinase and cleavage with CNBr were separated and purified by using reverse-plhase h.p.l.c. The amino acid sequence of each peptide was manually determined with the dimethylaminoazobenzene isothiocyanate/phenyl isothiocyanate double-coupling method. The primary structure of PIT-2 .beta.-lactamase was compared with those of two closely related enzymes, namely TEM-1 .beta.-lactamase and the .beta.-lactamase of Klebsiella pneumoniae strain LEN-1. The PIT-2 .beta.-lactamase amino acid sequence was strongly retained, with respectively 68% and 88% homology. Thus, PIT-2 enzyme could represent an evolutionary step between a chromosomally encoded .beta.-lactamase and the plasmid-mediated TEM .beta.-lactamases.