Purification and characterization of adenylate cyclase toxin from Bordetella pertussis.

Persistent Link:
http://hdl.handle.net/10150/184998
Title:
Purification and characterization of adenylate cyclase toxin from Bordetella pertussis.
Author:
Leusch, Mark Steven.
Issue Date:
1990
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Bordetella pertussis produces a number of virulence determinants believed to contribute to its survival in the host as well as to the pathogenesis of disease. One of these factors, adenylate cyclase toxin (ACT), has been implicated to penetrate human neutrophils and macrophages and abrogate their function by virtue of unregulated production of intracellular cAMP. In order to adequately study the nature of ACT and its role in pathogenesis, it is necessary to isolate the toxin from other virulence factors produced by the organism. Attempts by other investigators to purify ACT and maintain both its invasive and catalytic properties have not been successful. B. pertussis produces a cell associated ACT during mid-log phase of growth in Stainer-Scholte medium. Purification of ACT with both activities from urea extracted whole cells has been achieved by hydroxylapatite and calmodulin-sepharose chromatography. ACT is a single protein of 220 kd molecular weight with an isoelectric point of 7.0. The protein probably contains regions which are strongly hydrophobic. ACT has a specific activity of nearly 17,000 μM cAMP formed/min. An 850 ng sample of ACT induced over 1,400 pmoles cAMP/10⁶ S49 mouse lymphoma cells while 660 ng of ACT inhibited human neutrophil chemiluminescence by 65%.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Pertussis toxin -- Purification; Adenylate cyclase; Bordetella pertussis
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Microbiology and Immunology; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Friedman, Richard L.

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titlePurification and characterization of adenylate cyclase toxin from Bordetella pertussis.en_US
dc.creatorLeusch, Mark Steven.en_US
dc.contributor.authorLeusch, Mark Steven.en_US
dc.date.issued1990en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractBordetella pertussis produces a number of virulence determinants believed to contribute to its survival in the host as well as to the pathogenesis of disease. One of these factors, adenylate cyclase toxin (ACT), has been implicated to penetrate human neutrophils and macrophages and abrogate their function by virtue of unregulated production of intracellular cAMP. In order to adequately study the nature of ACT and its role in pathogenesis, it is necessary to isolate the toxin from other virulence factors produced by the organism. Attempts by other investigators to purify ACT and maintain both its invasive and catalytic properties have not been successful. B. pertussis produces a cell associated ACT during mid-log phase of growth in Stainer-Scholte medium. Purification of ACT with both activities from urea extracted whole cells has been achieved by hydroxylapatite and calmodulin-sepharose chromatography. ACT is a single protein of 220 kd molecular weight with an isoelectric point of 7.0. The protein probably contains regions which are strongly hydrophobic. ACT has a specific activity of nearly 17,000 μM cAMP formed/min. An 850 ng sample of ACT induced over 1,400 pmoles cAMP/10⁶ S49 mouse lymphoma cells while 660 ng of ACT inhibited human neutrophil chemiluminescence by 65%.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectPertussis toxin -- Purificationen_US
dc.subjectAdenylate cyclaseen_US
dc.subjectBordetella pertussisen_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineMicrobiology and Immunologyen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorFriedman, Richard L.en_US
dc.contributor.committeememberJoens, Lynen_US
dc.contributor.committeememberRyan, Kenen_US
dc.contributor.committeememberSterling, Chucken_US
dc.contributor.committeememberHubbard, Andreaen_US
dc.identifier.proquest9024509en_US
dc.identifier.oclc703889118en_US
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