Substrate specificity of the agonist-stimulated release of polyunsaturated fatty acids from vascular endothelial cells

Journal Article · Wed Nov 01 04:00:00 EST 1989 · Archives of Biochemistry and Biophysics; (USA)

DOI:https://doi.org/10.1016/0003-9861(89)90474-8· OSTI ID:5264767

Rosenthal, M D; Garcia, M C; Sprecher, H ^[1]

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1. Eastern Virginia Medical School, Norfolk (USA)

Stimulation of vascular endothelial cells with agonists such as histamine and thrombin results in release of arachidonic acid from membrane lipids and subsequent eicosanoid synthesis. As shown previously, the agonist-stimulated deacylation is specific for arachidonate, eicosapentaenoate, and 5,8,11-eicosatrienoate. This study has utilized radiolabeled fatty acids differing in chain length and position of double bonds to further elucidate the fatty acyl specificity of agonist-stimulated deacylation. Replicate wells of confluent human umbilical vein endothelial cells were incubated with 14C-labeled fatty acids and then challenged with histamine, thrombin, or the calcium ionophore A23187. Comparison of the results obtained with isomeric eicosatetraenoic fatty acids with initial double bonds at carbons 4, 5, or 6 indicated that the deacylation induced by all three agonists exhibited marked specificity for the cis-5 double bond. Lack of stringent chain length specificity was indicated by agonist-stimulated release of 5,8,11,14- tetraenoic fatty acids with 18, 19, 20, and 21 carbons. Release of 5,8,14-(14C)eicosatrienoate was two-to threefold that of 5,11,14-(14C)eicosatrienoate, thus indicating that the cis-8 double bond may also contribute to the stringent recognition by the agonist-sensitive phospholipase. The present study has also demonstrated that histamine, thrombin, and A23187 do not stimulate release of docosahexaenoate from endothelial cells.

• OSTI ID: 5264767
• Journal Information: Archives of Biochemistry and Biophysics; (USA), Journal Name: Archives of Biochemistry and Biophysics; (USA) Vol. 274:2; ISSN ABBIA; ISSN 0003-9861
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL TISSUES
ANIMALS
AZOLES
BODY
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CELL CULTURES
COAGULANTS
DRUGS
ENDOTHELIUM
ENZYMES
HEMATOLOGIC AGENTS
HEMOSTATICS
HETEROCYCLIC COMPOUNDS
HISTAMINE
HYDROLASES
IMIDAZOLES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MAN
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PEPTIDE HYDROLASES
PRIMATES
SECRETION
SERINE PROTEINASES
SPECIFICITY
STRUCTURE-ACTIVITY RELATIONSHIPS
SUBSTRATES
THROMBIN
TISSUES
TRACER TECHNIQUES
VERTEBRATES