Platelet activation initiates an upsurge in 18:2 and 20:4 lysophosphatidic acid (LPA) production. The biochemical pathway responsible for LPA production during blood clotting is not fully understood. We have purified a phospholipase A1 (PLA1) from thrombin-activated human platelets using sequential chromatographic steps followed by fluorophosphonate‑biotin affinity labeling and proteomics. We identified acyl‑protein thioesterase 1 (aka. lysophospholipase A1, accession code O75608) as a novel PLA1. Addition of this recombinant PLA1 significantly increased the production of sn‑2‑esterified polyunsaturated LPCs and the corresponding LPAs in plasma. We next examined the regioisomeric preference of lysophospholipase D/autotaxin (ATX), which is the subsequent step in LPA production. To prevent acylmigration regioisomers of oleyl‑sn‑glycero‑3‑phosphocholine (LPAF) were synthesized. ATX preferred the sn‑1 over the sn‑2 regioisomer of LPAF. We propose the following LPA production pathway in blood: 1) Activated platelets secrete PLA1. 2) PLA1generates a pool of sn‑2 lysophospholipids. 3) These newly generated sn‑2 lysophospholipids undergo acyl migration to yield sn‑1 lysophospholipids, which are the preferred substrates of ATX. 4) ATX cleaves the sn‑1 lysophospholipids to generate sn‑1 LPA species predominant with 18:2 and 20:4 fatty acids.
","abstract_html":"<p>Platelet activation initiates an upsurge in 18:2 and 20:4 lysophosphatidic acid (LPA) production. The biochemical pathway responsible for LPA production during blood clotting is not fully understood. We have purified a phospholipase A<sub>1</sub> (PLA<sub>1</sub>) from thrombin-activated human platelets using sequential chromatographic steps followed by fluorophosphonate‑biotin affinity labeling and proteomics. We identified acyl‑protein thioesterase 1 (aka. lysophospholipase A1, accession code O75608) as a novel PLA<sub>1</sub>. Addition of this recombinant PLA<sub>1</sub> significantly increased the production of <em>sn</em>‑<em>2</em>‑esterified polyunsaturated LPCs and the corresponding LPAs in plasma. We next examined the regioisomeric preference of lysophospholipase D/autotaxin (ATX), which is the subsequent step in LPA production. To prevent acylmigration regioisomers of oleyl‑<em>sn</em>‑glycero‑3‑phosphocholine (LPAF) were synthesized. ATX preferred the <em>sn</em>‑<em>1</em> over the <em>sn</em>‑<em>2</em> regioisomer of LPAF. We propose the following LPA production pathway in blood: 1) Activated platelets secrete PLA<sub>1</sub>. 2) PLA<sub>1</sub>generates a pool of <em>sn‑2 </em>lysophospholipids. 3) These newly generated <em>sn‑2 </em>lysophospholipids undergo acyl migration to yield <em>sn‑1 </em>lysophospholipids, which are the preferred substrates of ATX. 4) ATX cleaves the <em>sn‑1 </em>lysophospholipids to generate <em>sn‑1 </em>LPA species predominant with 18:2 and 20:4 fatty acids.</p>","abstract_has_math":false,"creators":["Bolen, Alyssa Lynn Jefferson"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation","degree_discipline":"Biomedical Sciences","degree_department":null,"school":null,"contributors":["Gabor J. Tigyi, M.D., Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-01T07:00:00Z","date_published":"2011-05-01T07:00:00Z","updated_at":"2026-07-24T05:00:02Z","subjects":["Autotaxin","Lysophosphatidic acid","Phospholipase A1","Medical Cell Biology","Medical Molecular Biology","Medical Sciences","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://dc.uthsc.edu/dissertations/27","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gabor J. Tigyi, M.D., Ph.D."]},{"key":"dc:creator","label":"Author","values":["Bolen, Alyssa Lynn Jefferson"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2016-05-17T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biomedical Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Autotaxin","Lysophosphatidic acid","Phospholipase A1","Medical Cell Biology","Medical Molecular Biology","Medical Sciences","Medicine and Health Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://dc.uthsc.edu/dissertations/27"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Platelet activation initiates an upsurge in 18:2 and 20:4 lysophosphatidic acid (LPA) production. The biochemical pathway responsible for LPA production during blood clotting is not fully understood. We have purified a phospholipase A1 (PLA1) from thrombin-activated human platelets using sequential chromatographic steps followed by fluorophosphonate‑biotin affinity labeling and proteomics. We identified acyl‑protein thioesterase 1 (aka. lysophospholipase A1, accession code O75608) as a novel PLA1. Addition of this recombinant PLA1 significantly increased the production of sn‑2‑esterified polyunsaturated LPCs and the corresponding LPAs in plasma. We next examined the regioisomeric preference of lysophospholipase D/autotaxin (ATX), which is the subsequent step in LPA production. To prevent acylmigration regioisomers of oleyl‑sn‑glycero‑3‑phosphocholine (LPAF) were synthesized. ATX preferred the sn‑1 over the sn‑2 regioisomer of LPAF. We propose the following LPA production pathway in blood: 1) Activated platelets secrete PLA1. 2) PLA1generates a pool of sn‑2 lysophospholipids. 3) These newly generated sn‑2 lysophospholipids undergo acyl migration to yield sn‑1 lysophospholipids, which are the preferred substrates of ATX. 4) ATX cleaves the sn‑1 lysophospholipids to generate sn‑1 LPA species predominant with 18:2 and 20:4 fatty acids.
"]},{"key":"dc:title","label":"Title","values":["The Biochemical Pathway Leading to Lpa Generation Upon Blood Coagulation"]}]}],"canonical_facts":{"dc:contributor":["Gabor J. Tigyi, M.D., Ph.D."],"dc:creator":["Bolen, Alyssa Lynn Jefferson"],"dc:date.available":["2016-05-17T07:00:00Z"],"dc:description.abstract":["Platelet activation initiates an upsurge in 18:2 and 20:4 lysophosphatidic acid (LPA) production. The biochemical pathway responsible for LPA production during blood clotting is not fully understood. We have purified a phospholipase A1 (PLA1) from thrombin-activated human platelets using sequential chromatographic steps followed by fluorophosphonate‑biotin affinity labeling and proteomics. We identified acyl‑protein thioesterase 1 (aka. lysophospholipase A1, accession code O75608) as a novel PLA1. Addition of this recombinant PLA1 significantly increased the production of sn‑2‑esterified polyunsaturated LPCs and the corresponding LPAs in plasma. We next examined the regioisomeric preference of lysophospholipase D/autotaxin (ATX), which is the subsequent step in LPA production. To prevent acylmigration regioisomers of oleyl‑sn‑glycero‑3‑phosphocholine (LPAF) were synthesized. ATX preferred the sn‑1 over the sn‑2 regioisomer of LPAF. We propose the following LPA production pathway in blood: 1) Activated platelets secrete PLA1. 2) PLA1generates a pool of sn‑2 lysophospholipids. 3) These newly generated sn‑2 lysophospholipids undergo acyl migration to yield sn‑1 lysophospholipids, which are the preferred substrates of ATX. 4) ATX cleaves the sn‑1 lysophospholipids to generate sn‑1 LPA species predominant with 18:2 and 20:4 fatty acids.
"],"dc:identifier":["https://dc.uthsc.edu/dissertations/27"],"dc:subject":["Autotaxin","Lysophosphatidic acid","Phospholipase A1","Medical Cell Biology","Medical Molecular Biology","Medical Sciences","Medicine and Health Sciences"],"dc:title":["The Biochemical Pathway Leading to Lpa Generation Upon Blood Coagulation"],"thesis:degree_discipline":["Biomedical Sciences"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:00:02Z"}