Identification and characterization of a thermolabile antigen (TLAa, enolase) in Saccharomyces cerevisiae.
Five kinds of proteins of Saccharomyces cerevisiae were recently purified to a homogeneous state and identified as yeast cell surface antigens (TLAa, TLAb, TLAc, TLAd, and TLAe), but their physiological functions remained uncertain. In this paper, TLAa was identified as a yeast enolase (EC 4.2.1.11) from the following evidence: (1) molecular weights and amino acid compositions of both proteins were similar, (2) the N-terminal 22 amino acid sequences of both proteins were the same (3), TLAa had enolase activity, and (4) the authentic yeast enolase gave a positive reaction with anti-TLAa serum in the Ouchterlony immunodiffusion test and the immunoblotting test. Two isoproteins of TLAa (enolase) were separated by non-denatured polyacrylamide gel electrophoresis and detected by immunoblotting with anti-TLAa serum. The contents of the two isoproteins varied depending on the following culture conditions: glucose starvation, growth in the presence of non-fermentable carbon sources, and growth in media containing sodium chloride and 2-deoxyglucose. The contents did not vary, however, with heat shock treatment or with growth in media containing sodium azide, tunicamycin, or sorbitol. These results showed that TLAa was a cytoplasmic antigen, and that its synthesis was regulated by some environmental stresses.
Department of Bioresources, Faculty of Agriculture, Ehime University, Japan.
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Watanabe
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Ikeuchi
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Tamai
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000946:Antigens, Fungal / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000737:chemistry; D015151:Immunoblotting; D005779:Immunodiffusion; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010751:Phosphopyruvate Hydratase / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000254:growth & development / Q000276:immunology*
D000596:Amino Acids; D000946:Antigens, Fungal; D000954:Antigens, Surface; D006360:Heat-Shock Proteins; D010751:Phosphopyruvate Hydratase