Purification and sequencing of the active site tryptic peptide from penicillin-binding protein 1b of Escherichia coli
This paper reports the sequence of the active site peptide of penicillin-binding protein 1b from Escherichia coli. Purified penicillin-binding protein 1b was labeled with (/sup 14/C)penicillin G, digested with trypsin, and partially purified by gel filtration. Upon further purification by high-pressure liquid chromatography, two radioactive peaks were observed, and the major peak, representing over 75% of the applied radioactivity, was submitted to amino acid analysis and sequencing. The sequence Ser-Ile-Gly-Ser-Leu-Ala-Lys was obtained. The active site nucleophile was identified by digesting the purified peptide with aminopeptidase M and separating the radioactive products on high-pressure liquid chromatography. Amino acid analysis confirmed that the serine residue in the middle of the sequence was covalently bonded to the (/sup 14/C)penicilloyl moiety. A comparison of this sequence to active site sequences of other penicillin-binding proteins and beta-lactamases is presented.
- Research Organization:
- Harvard Univ., Cambridge, MA
- OSTI ID:
- 6171278
- Journal Information:
- Biochemistry; (United States), Vol. 14
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PEPTIDES
AMINO ACID SEQUENCE
PURIFICATION
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
CARBON 14 COMPOUNDS
CARBOXYPEPTIDASES
ESCHERICHIA COLI
LIQUID COLUMN CHROMATOGRAPHY
PENICILLIN
TRACER TECHNIQUES
TRYPSIN
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BACTERIA
CARBOXYLIC ACIDS
CHROMATOGRAPHY
DRUGS
ENZYMES
HYDROLASES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MICROORGANISMS
MOLECULAR STRUCTURE
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
PROTEINS
REACTION KINETICS
SEPARATION PROCESSES
SERINE PROTEINASES
550201* - Biochemistry- Tracer Techniques