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Title: Lymphocyte receptors for pertussis toxin

Abstract

We have investigated human T-lymphocyte receptors for pertussis toxin by affinity isolation and photoaffinity labeling procedures. T lymphocytes were obtained from peripheral human blood, surface iodinated, and solubilized in Triton X-100. The iodinated mixture was then passed through pertussis toxin-agarose, and the fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Autoradiography of the fixed, dried gels revealed several bands in the pertussis toxin-bound fraction that were not observed in fractions obtained from histone or fetuin-agarose. Further investigations employed a photoaffinity labeling reagent, sulfosuccinimidyl 2-(p-azido-salicylamido)-1,3'-dithiopropionate, to identify pertussis toxin receptors in freshly isolated peripheral blood monocytic cells, T lymphocytes, and Jurkat cells. In all three cell systems, the pertussis toxin affinity probe specifically labeled a single protein species with an apparent molecular weight of 70,000 that was not observed when the procedure was performed in the presence of excess unmodified pertussis toxin. A protein comparable in molecular weight to the one detected by the photoaffinity labeling technique was also observed among the species that bound to pertussis toxin-agarose. The results suggest that pertussis toxin may bind to a 70,000-Da receptor in human T lymphocytes.

Authors:
;  [1]
  1. (Univ. of Alberta, Edmonton (Canada))
Publication Date:
OSTI Identifier:
6171258
Resource Type:
Journal Article
Resource Relation:
Journal Name: Infection and Immunity; (USA); Journal Volume: 58:12
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; LYMPHOCYTES; RECEPTORS; LABELLING; TOXINS; BIOCHEMICAL REACTION KINETICS; AFFINITY; AUTORADIOGRAPHY; ELECTROPHORESIS; IODINE ISOTOPES; MAN; MOLECULAR WEIGHT; REAGENTS; ANIMAL CELLS; ANIMALS; ANTIGENS; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CONNECTIVE TISSUE CELLS; ISOTOPES; KINETICS; LEUKOCYTES; MAMMALS; MATERIALS; MEMBRANE PROTEINS; ORGANIC COMPOUNDS; PRIMATES; PROTEINS; REACTION KINETICS; SOMATIC CELLS; TOXIC MATERIALS; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Clark, C.G., and Armstrong, G.D. Lymphocyte receptors for pertussis toxin. United States: N. p., 1990. Web.
Clark, C.G., & Armstrong, G.D. Lymphocyte receptors for pertussis toxin. United States.
Clark, C.G., and Armstrong, G.D. 1990. "Lymphocyte receptors for pertussis toxin". United States. doi:.
@article{osti_6171258,
title = {Lymphocyte receptors for pertussis toxin},
author = {Clark, C.G. and Armstrong, G.D.},
abstractNote = {We have investigated human T-lymphocyte receptors for pertussis toxin by affinity isolation and photoaffinity labeling procedures. T lymphocytes were obtained from peripheral human blood, surface iodinated, and solubilized in Triton X-100. The iodinated mixture was then passed through pertussis toxin-agarose, and the fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Autoradiography of the fixed, dried gels revealed several bands in the pertussis toxin-bound fraction that were not observed in fractions obtained from histone or fetuin-agarose. Further investigations employed a photoaffinity labeling reagent, sulfosuccinimidyl 2-(p-azido-salicylamido)-1,3'-dithiopropionate, to identify pertussis toxin receptors in freshly isolated peripheral blood monocytic cells, T lymphocytes, and Jurkat cells. In all three cell systems, the pertussis toxin affinity probe specifically labeled a single protein species with an apparent molecular weight of 70,000 that was not observed when the procedure was performed in the presence of excess unmodified pertussis toxin. A protein comparable in molecular weight to the one detected by the photoaffinity labeling technique was also observed among the species that bound to pertussis toxin-agarose. The results suggest that pertussis toxin may bind to a 70,000-Da receptor in human T lymphocytes.},
doi = {},
journal = {Infection and Immunity; (USA)},
number = ,
volume = 58:12,
place = {United States},
year = 1990,
month =
}
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