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Identification of receptor regulatory proteins, membrane glycoproteins, and functional characteristics of adenylate cyclase in vesicles derived from the human neutrophil

Lad, P. M. and Glovsky, M. M. and Richards, J. H. and Learn, D. B. and Reisinger, D. M. and Smiley, P. A. (1984) Identification of receptor regulatory proteins, membrane glycoproteins, and functional characteristics of adenylate cyclase in vesicles derived from the human neutrophil. Molecular Immunology, 21 (7). pp. 627-639. ISSN 0161-5890. https://resolver.caltech.edu/CaltechAUTHORS:20150505-075621107

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Abstract

Human neutrophils were disrupted by brief sonication under conditions which preserve the hormone sensitivity of adenylate cyclase and yield minimal granule lysis. Fractions enriched in adenylate cyclase were analysed for hormonal and guanine nucleotide regulation of the enzyme as well as structural proteins. Adenylate cyclase was activated by PGE_1 and isoproterenol in a GTP-dependent fashion, while f-met-leu-phe and C5a gave no stimulation. Cholera toxin treatment, which specifically modifies cyclase-related GTP-binding proteins, caused a dose-dependent enhancement of GTP activation, in which GTP alone activated maximally and PGE1 was without further effect. The following proteins were detected in the cyclase-containing vesicles: (a) a 42 K mol. wt protein labeled selectively by cholera toxin; (b) protein subunits observed in SDS gels at 214, 165, 105 and 47 K, of which the 47 K band was the most prominent and comigrated with actin; (c) prominent lectin-binding activities at 165 K (concanavalin A and wheat germ agglutinin) as well as at 100 K (wheat germ agglutinin); and (d) a set of proteins and lectin-binding activities in fractions containing β-glucuronidase activity distinct from adenylate cyclase containing vesicles. The identification of receptor-controlled cyclase, GTP-binding regulatory proteins, cytoskeletal elements and unique lectin-binding activities in a single vesicle preparation should contribute to an understanding of receptor-mediated control of neutrophil function.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0161-5890(84)90048-8DOIArticle
http://www.sciencedirect.com/science/article/pii/0161589084900488PublisherArticle
Additional Information:© 1984 Pergamon Press Ltd. First received 9 November 1983; accepted in revised form 17 February 1984. This work was supported by grant No. 17 from the Department of Education and Research, Kaiser-Permanente, Southern California Permanente Medical Group and in part by a Biomedical Research Support Grant RR 05521-19 from the Kaiser Foundation Research Institute. We thank Aline Alenty for advice and assistance in the preparation of neutrophils and in neutrophil function tests. We are also grateful to Lauren Rosenthal and Bill Frankeberger for secretarial assistance. Publication No. 72 from the Kaiser Regional Research Laboratory.
Funders:
Funding AgencyGrant Number
Kaiser-Permanente17
Kaiser Foundation Research InstituteRR 05521-19
Issue or Number:7
Record Number:CaltechAUTHORS:20150505-075621107
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150505-075621107
Official Citation:P.M. Lad, M.M. Glovsky, J.H. Richards, D.B. Learn, D.M. Reisinger, P.A. Smiley, Identification of receptor regulatory proteins, membrane glycoproteins, and functional characteristics of adenylate cyclase in vesicles derived from the human neutrophil, Molecular Immunology, Volume 21, Issue 7, July 1984, Pages 627-639, ISSN 0161-5890, http://dx.doi.org/10.1016/0161-5890(84)90048-8. (http://www.sciencedirect.com/science/article/pii/0161589084900488)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:57214
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:06 May 2015 16:42
Last Modified:03 Oct 2019 08:22

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