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Concurrent assessment of inner and outer membrane permeabilization and bacteriolysis in E. coli by multiple-wavelength spectrophotometry.

Abstract：

:We developed a dual wavelength spectrophotometric assay that permitted beta-lactamase and beta-galactosidase activities to be measured concurrently in a single sample. We also constructed a target cell, E. coli ML-35p, that was substantially cryptic for its periplasmic beta-lactamase and cytoplasmic beta-galactosidase unless outer membrane (beta-lactamase) or inner membrane (beta-galactosidase) permeabilization occurred. By applying the spectrophotometric assay to whole target cells, we could ascertain the kinetics of inner and outer membrane permeabilization by biological agents, including serum, polymyxin B and mellitin. By monitoring the reactions at an additional wavelength, we could also follow the kinetics of serum-mediated bacteriolysis. These experiments illustrate the principle of multiple wavelength spectrophotometry and provide examples of its use to monitor and dissect the action of biological agents on a gram-negative bacterium.

journal_name

J Immunol Methods

journal_title

Journal of immunological methods

authors

Lehrer RI,Barton A,Ganz T

doi

10.1016/0022-1759(88)90414-0

subject

Has Abstract

pub_date

1988-04-06 00:00:00

pages

153-8

issue

1-2

eissn

0022-1759

issn

1872-7905

pii

0022-1759(88)90414-0

journal_volume

108

pub_type

杂志文章

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