Isolation and characterization of hemagglutinins from Nereis virens: A polychaetous annelid.
Item
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Title
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Isolation and characterization of hemagglutinins from Nereis virens: A polychaetous annelid.
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Identifier
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AAI9000040
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identifier
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9000040
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Creator
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Lai, Pi-shiang.
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Contributor
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Adviser: Charlotte S. Russell
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Date
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1989
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Language
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English
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Publisher
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City University of New York.
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Subject
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Chemistry, Biochemistry
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Abstract
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Colemic fluid was fractionated into low (LMW) and high (HMW) molecular weight and lipid extract (CM) agglutinins.;LMW contains four (I-IV) heat-sensitive and Ca{dollar}\sp{lcub}++{rcub}{dollar}/Mg{dollar}\sp{lcub}++{rcub}{dollar}-independent glycoproteins with specific erythrocyte (RBC) and inhibition profiles and MW 20, 20, 19, 24 kd respectively. I-IV are inhibited by fetuin {dollar}\geq{dollar} bovine submaxillary mucin (BSM) {dollar}>{dollar} thyroglobulin {dollar}>{dollar} yeast invertase {dollar}\geq{dollar} mannan. I-IV inhibited by mannose-1-phosphate. Immunoelectrophoresis shows precipitin lines which coincide with activity, protein and carbohydrate stains. LMW I is catonic, LMW II, uncharged and LMW III & IV anionic at pH 8.3. LMW protects mice from proliferation of Ehrlich ascites tumor cells (EATC).;HMW is a lipid-associated sulfated proteoglycan which agglutinates rabbit and rat RBC. Activity is inhibited by fetuin {dollar}>{dollar} BSM {dollar}>{dollar} DsOSM {dollar}>{dollar} OSM. In immunoelectrophoresis (at pH 8.3), (a) HMW gives two precipitin lines which coincide with the region of activity, which also stains for lipid, sugar and protein. (b) HHMW and DL-HHMW do not give any precipitin line but the region of activity is the same as that of HMW. DL-HHMW contains protein (12%), hexose (30%), hexuronate (9%), hexosamine (8%), glycosaminoglycan (GAG, at least 22%) and sulfate (7%). DL-HHMW gives a metachromatic reaction with dimethymethylene blue. The infrared spectrum of DL-HHMW shows a hexose-6-sulfate ester. Chondroitinase ABC, keratanase and heparinase II do not change the activity and GAG content. HMW delays the growth of EATC in mice.;CM is heat-insensitive and Ca{dollar}\sp{lcub}++{rcub}{dollar}/Mg{dollar}\sp{lcub}++{rcub}{dollar}-dependent and agglutinates rat {dollar}>{dollar} rabbit {dollar}>{dollar} green monkey {dollar}>{dollar} rhesus monkey = human (O) = guinea pig {dollar}\gg{dollar} chicken RBC. Agglutination is inhibited by BSM {dollar}>{dollar} HGM {dollar}\geq{dollar} fetuin. In immunoelectrophoresis (at pH 8.3), CM does not gives any precipitin line and the region of activity extends along the length of the gel. CM was resolved by 2D TLC on silica and studied with group specific reagents and IR. One of the active spots contains phosphate, -NH{dollar}\sb2{dollar}, fatty acid ester and vicinal-OH and migrates like DOPE. CM retards proliferation of EATC in mice.;Contradictory reports in the literature about the hemagglutinating activity of DOPE are partially-explained. DOPE itself is not an agglutinin but oxidized DOPE is.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
