Molecular characterization of thermoinduced immunogenic proteins Q1p42 and Hsp15 of Leptospira interrogans.
Leptospira interrogans is a mammalian pathogen which must adapt to a range of new environmental conditions including temperature change when it infects new hosts. In vitro studies of organisms cultured at 30 degrees C and shifted to 37 degrees C for 5 to 7 days have confirmed that synthesis of several proteins involved in equine infection is regulated in response to temperature change (J. E. Nally, J. F. Timoney, and B. Stevenson, Infect. Immun. 69:400-404, 2001). In order to specifically identify antigenic proteins upregulated at 37 degrees C, groups of three ponies were immunized with organisms shifted to 37 degrees C for 5 to 7 days or with organisms maintained at 30 degrees C. A lambda ZAP II genomic DNA library was screened with the pool of antisera to organisms shifted to 37 degrees C. Clones reactive with this pool but unreactive with the pool of pony antisera to organisms cultured at 30 degrees C were selected for further analysis. Sequence analysis of the first two clones identified open reading frames for proteins designated Qlp42 and Hsp15. Qlp42 is predicted to be an outer membrane lipoprotein. Its synthesis was upregulated when cultures were shifted from 30 to 37 degrees C and downregulated when cultures were shifted from 37 to 30 degrees C. Although the predicted molecular mass of Qlp42 is 39.8 kDa for the mature protein, Qlp42-specific equine antiserum was reactive with two bands of 30 and 29.5 kDa. Hsp15 is a stress response protein and a member of the Hsp20/alpha-crystallin family. PCR detected homologues of qlp42 and hsp15 in pathogenic serovars of L. interrogans but not in the nonpathogenic Leptospira biflexa. Enzyme-linked immunosorbent assays of antibody in convalescent sera from mares naturally infected with L. interrogans suggest that Qlp42 is expressed during leptospiral infection.
Nally
J E
JE
Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 40546-0099, USA.
Artiushin
S
S
Timoney
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003289:Convalescence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology*; D029968:Escherichia coli Proteins; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D015698:Genomic Library; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology*; D006734:Horse Diseases / Q000097:blood / Q000276:immunology; D006736:Horses; D006358:Hot Temperature; D007921:Leptospira interrogans / Q000235:genetics / Q000276:immunology*; D007922:Leptospirosis / Q000097:blood / Q000276:immunology / Q000662:veterinary; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015854:Up-Regulation
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D008074:Lipoproteins; C440648:Qlp42 protein, Leptospira interrogans; C116841:hslR protein, E coli