Distinct immunoreactivities suggest the existence of potential tissue variants in rat lipoprotein lipase.
Lipoprotein lipases (LPL) isolated from rat cardiac muscle and bovine milk were each used as immunogens to produce polyclonal anti-LPL sera and two anti-LPL monoclonal antibodies. The immunological reactivities of these antibody sources with LPL purified from rat cardiac muscle, lung, adipose tissue, mammary gland and skeletal muscle were compared by an e.l.i.s.a. and by Western blotting. Differences between the immunoreactivities of LPL from the distinct tissue sources were revealed in both systems. A synthetic peptide with a sequence corresponding to the heparin-binding site of LPL (Ser-Arg-Thr-Asn-Thr-Lys-Val-Ser-Arg-Ile-Thr-Gly-Leu) was produced and used as an immunogen. The antiserum produced against the synthetic peptide was found to bind specifically to the region of the heparin-binding site, as determined by use of a competition e.l.i.s.a. In use against the five tissue LPL preparations, this antiserum revealed only minor variations between the tissue sources, compared with the hierarchy of reactivity observed when antibodies raised against the whole molecule were used. In combination with the outcome of previous studies on some of the physical properties of these preparations [Soteriou and Cryer (1993) Int. J. Biochem. 25, 1483-1490], the observations reported here on the distinct immunoreactivities exhibited by LPL prepared from the different tissue sources of a single species indicate the necessity to characterize fully the nature of these differences.
Soteriou
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Department of Biochemistry, University of Wales College of Cardiff, U.K.
Cryer
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D000273:Adipose Tissue / Q000201:enzymology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay; D014644:Genetic Variation; D006493:Heparin / Q000378:metabolism; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008071:Lipoprotein Lipase / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D008168:Lung / Q000201:enzymology; D008321:Mammary Glands, Animal / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D009928:Organ Specificity; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D000911:Antibodies, Monoclonal; D009842:Oligopeptides; D006493:Heparin; D008071:Lipoprotein Lipase