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2008-12-22Zeitschriftenartikel DOI: 10.18452/9444
Thrombin induces broad spectrum proteolysis in human serum samples
dc.contributor.authorO'Mullan, Patrick
dc.contributor.authorCraft, David
dc.contributor.authorYi, Jizu
dc.contributor.authorGelfand, Craig A.
dc.date.accessioned2017-06-17T01:06:19Z
dc.date.available2017-06-17T01:06:19Z
dc.date.created2009-06-26
dc.date.issued2008-12-22
dc.identifier.issn1437-4331
dc.identifier.urihttp://edoc.hu-berlin.de/18452/10096
dc.description.abstractBackground: During clotting, α thrombin cleaves fibrinogen releasing fibrinopeptide A (FPA). FPA is easily identified in serum using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Using MALDI-TOF MS, we observed multiple, progressively shorter fragments of serum FPA. Following ambient incubation of serum, variations in the content of FPA fragments occur over time. Denaturation of α thrombin by heating the serum sample appears to minimize this variation. These observations suggest that intrinsic proteolytic and peptidolytic activity is elevated in serum and perhaps originates from the coagulation cascade enzymes themselves, especially α thrombin. // Methods: Extrinsic addition of α thrombin to a subset (3–30 kDa) of plasma proteins was carried out to induce proteolysis and to examine the resultant peptides to reveal α thrombin susceptible parent proteins. One of these identified proteins, hemopexin, was directly digested by α thrombin and the peptides examined to confirm the observations from the initial plasma protein digestion. // Results: Extrinsic addition of α thrombin to a subset (3–30 kDa) of plasma proteins results in wide-spread digestion of proteins unrelated to coagulation, revealing a substrate range encompassing more than fibrinogen. Direct digestion of one of these proteins, hemopexin, by α thrombin confirms these observations. // Conclusions: The resulting peptides indicate broad tolerance beyond the consensus R-G cleavage site of fibrinogen; in fact, there appears to be no bias for the amino acid following the R/K residue. These data support our hypothesis that the enzymatic activities inherent to coagulation, or at least to thrombin, contribute to destabilization of the protein and peptide content of serum.eng
dc.language.isoeng
dc.publisherKooperation de Gruyter
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/
dc.subjectfibrinopeptideeng
dc.subjectproteolysiseng
dc.subjectthrombineng
dc.subject.ddc540 Chemie und zugeordnete Wissenschaften
dc.subject.ddc610 Medizin und Gesundheit
dc.titleThrombin induces broad spectrum proteolysis in human serum samples
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-10098712
dc.identifier.doihttp://dx.doi.org/10.18452/9444
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-year2009
dc.description.versionPeer Reviewed
dcterms.bibliographicCitation.doi10.1515/CCLM.2009.003
dcterms.bibliographicCitation.journaltitleClinical chemistry and laboratory medicine
dcterms.bibliographicCitation.volume47
dcterms.bibliographicCitation.issue6
dcterms.bibliographicCitation.pagestart685
dcterms.bibliographicCitation.pageend693

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