مورد إلكتروني
Covalent binding of arachidonic acid metabolites to human platelet proteins. Identification of prostaglandin H synthase as one of the modified substrates.
| العنوان: | Covalent binding of arachidonic acid metabolites to human platelet proteins. Identification of prostaglandin H synthase as one of the modified substrates. |
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| المصدر: | The Journal of biological chemistry, 265 (9 |
| بيانات النشر: | 1990-03 |
| تفاصيل مُضافة: | Lecomte, Marc Lecocq, Raymond Dumont, Jacques Emile Boeynaems, Jean-Marie |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | The covalent modification of proteins by metabolites of arachidonic acid (AA) was investigated in human platelets. Following incubation of washed human platelets with radiolabeled AA, ethanol precipitation of the proteins, and lipid extraction by organic solvents, a small fraction of the radioactivity added (0.3%) was tightly bound to the protein pellet. A dozen labeled protein bands were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Exhaustive hydrolysis of platelet proteins by proteases released an amphipathic radiolabeled material which had a chromatographic behavior similar to that of a known peptidolipid, leukotriene C4. These findings suggest a covalent nature for the observed binding. This binding was specific for AA since palmitate, myristate, or linoleate did not bind to a significant extent. It involved products of both cyclooxygenase and lipoxygenase pathways: it was indeed inhibited to a greater extent by eicosatetraynoic acid than by indomethacin. The protein-associated radioactivity was increased by the thromboxane synthase inhibitor dazoxiben. Indomethacin completely abolished this increase in binding, which could not be reproduced by exogenous prostaglandin (PG) E2, F2 alpha, or D2, and might thus involve PGG2 and/or PGH2. Diamide, an agent known to inhibit the reduction of 12-hydroperoxyeicosatetraenoic acid in platelets, produced an increase of the covalent binding, which was abolished by eicosatetraynoic acid but not by indomethacin: this suggests that the lipoxygenase product bound was 12-hydroperoxyeicosatetraenoic acid or a by-product. Dazoxiben and diamide produced distinct patterns of protein labeling after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. One labeled band had a Mr of 70,000 as the PGH synthase monomer. Addition of AA at 17 microM enhanced the labeling of this band, while 100 microM was inhibitory. Labeling of this band was also induced by thrombin in prelabeled platelets. Two monoclonal anti Journal Article Research Support, Non-U.S. Gov't info:eu-repo/semantics/published |
| مصطلحات الفهرس: | Sciences bio-médicales et agricoles, Animals, Arachidonic Acids -- blood, Blood Platelets -- metabolism, Blood Proteins -- isolation & purification, Blood Proteins -- metabolism, Electrophoresis, Polyacrylamide Gel, Humans, Kinetics, Male, Molecular Weight, Prostaglandin-Endoperoxide Synthases -- blood, Prostaglandin-Endoperoxide Synthases -- isolation & purification, Prostaglandin-Endoperoxide Synthases -- metabolism, Protein Binding, Seminal Vesicles -- enzymology, Sheep, info:eu-repo/semantics/article, info:ulb-repo/semantics/articlePeerReview, info:ulb-repo/semantics/openurl/article |
| URL: | |
| الإتاحة: | Open access content. Open access content 1 full-text file(s): info:eu-repo/semantics/openAccess |
| ملاحظة: | 1 full-text file(s): application/pdf English |
| أرقام أخرى: | EQY oai:dipot.ulb.ac.be:2013/50210 uri/info:pmid/2108168 uri/info:scp/0025234871 local/VX-006810 1363793304 |
| المصدر المساهم: | UNIVERSITE LIBRE DE BRUXELLES From OAIster®, provided by the OCLC Cooperative. |
| رقم الأكسشن: | edsoai.on1363793304 |
| قاعدة البيانات: | OAIster |
| الوصف غير متاح. |