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PMID: 4630162 Published · ppublish English Journal Article

Isolation by covalent affinity chromatography of the penicillin-binding components from membranes of Bacillus subtilis.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 69 ·No. 12 ·1972-12-00 ·Pages 3751-5

Blumberg PM, Strominger JL

Abstract

An affinity chromatography technique was developed to isolate the five penicillin-binding components present in Bacillus subtilis membranes. The proteins were solubilized by the detergent Nonidet P-40, bound covalently to penicillin-substituted Sepharose, and subsequently eluted from the matrix with neutral hydroxylamine, which cleaves the penicilloyl-enzyme bond. Penicillin binding-component V, the D-alanine carboxypeptidase, makes up 1% of the total membrane protein. A modification of the above procedure enabled this enzyme to be obtained from the membrane in pure form in a single step with 50% overall recovery of enzymatic activity.

MeSH Terms
Alanine Bacillus subtilis/analysis,enzymology Bacterial Proteins/isolation & purification,metabolism Binding Sites Carbon Isotopes Carboxypeptidases/isolation & purification Cell Membrane/analysis Chromatography Chromatography, Affinity Electrophoresis, Polyacrylamide Gel Penicillin G/metabolism Protein Binding
Chemicals
Bacterial Proteins Carbon Isotopes Carboxypeptidases Alanine Penicillin G
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Blumberg P M
Strominger J L
References (25)
25 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.69.12.3751
Published
1972-12-00
Pages
3751-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC389864
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.69.12.3751 PMC Full Text View journal details
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