Please use this identifier to cite or link to this item: https://ahro.austin.org.au/austinjspui/handle/1/13106
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dc.contributor.authorSmyth, Mark Jen
dc.contributor.authorMcGuire, M Jen
dc.contributor.authorThia, K Yen
dc.date.accessioned2015-05-16T02:53:21Z
dc.date.available2015-05-16T02:53:21Z
dc.date.issued1995-06-15en
dc.identifier.citationJournal of Immunology (baltimore, Md. : 1950); 154(12): 6299-305en
dc.identifier.govdoc7759868en
dc.identifier.otherPUBMEDen
dc.identifier.urihttps://ahro.austin.org.au/austinjspui/handle/1/13106en
dc.description.abstractHuman granzyme B (hGrzB) is the key member of a family of granule serine proteases (granzymes) that participate in target cell death inflicted by cytotoxic lymphocytes. The proenzyme activation peptide predicted from the cDNA encoding hGrzB is composed of two residues. We have devised a PCR strategy to delete this activation dipeptide within hGrzB and express active recombinant hGrzB in mammalian COS cells. Lysates of COS cells transfected with modified hGrzB cDNA were able to hydrolyze tert-butyloxycarbonyl-Ala-Ala-Asp-thiobenzyl ester (Boc-Ala-Ala-Asp-SBzl), whereas lysates transfected with unmodified hGrzB cDNA were inactive. Accordingly, active recombinant hGrzB displayed no significant activities toward substrates containing Met-, Lys-, or Phe- at P1. It has been suggested that the mechanism by which the amino-terminal dipeptide is normally cleaved to generate active GrzB involves dipeptidyl peptidase I (DPPI). Our studies demonstrated that lysates of COS cells cotransfected with unmodified hGrzB cDNA (inactive) and rat DPPI cDNA were able to hydrolyze Boc-Ala-Ala-Asp-SBzl. Similarly, lysates of COS cells transfected with unmodified hGrzB cDNA, and devoid of Asp-ase activity, were also activated upon the addition of bovine spleen DPPI in a pH and time-dependent fashion. These results suggest that the activation dipeptide, and more particularly DPPI, may play and important role in the normal post-translational processing and activation of hGrzB in cytotoxic lymphocytes.en
dc.language.isoenen
dc.subject.otherAnimalsen
dc.subject.otherBase Sequenceen
dc.subject.otherCathepsin Cen
dc.subject.otherCell Lineen
dc.subject.otherDNA Primers.geneticsen
dc.subject.otherDNA, Complementary.geneticsen
dc.subject.otherDipeptidyl-Peptidases and Tripeptidyl-Peptidases.metabolismen
dc.subject.otherEnzyme Activationen
dc.subject.otherGene Expressionen
dc.subject.otherGranzymesen
dc.subject.otherHumansen
dc.subject.otherMolecular Sequence Dataen
dc.subject.otherProtein Processing, Post-Translationalen
dc.subject.otherRecombinant Proteins.genetics.metabolismen
dc.subject.otherSerine Endopeptidases.genetics.metabolismen
dc.subject.otherT-Lymphocytes, Cytotoxic.enzymology.immunologyen
dc.subject.otherTransfectionen
dc.titleExpression of recombinant human granzyme B. A processing and activation role for dipeptidyl peptidase I.en
dc.typeJournal Articleen
dc.identifier.journaltitleJournal of Immunology (Baltimore, Md. : 1950)en
dc.identifier.affiliationCellular Cytotoxicity Laboratory, Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australiaen
dc.description.pages6299-305en
dc.relation.urlhttps://pubmed.ncbi.nlm.nih.gov/7759868en
dc.type.austinJournal Articleen
item.openairetypeJournal Article-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.grantfulltextnone-
item.fulltextNo Fulltext-
item.cerifentitytypePublications-
item.languageiso639-1en-
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