The 18-kDa cytoplasmic protein of Brucella species -: an antigen useful for diagnosis -: is a lumazine synthase

被引:49
作者
Goldbaum, FA
Velikovsky, CA
Baldi, PC
Mörtl, S
Bacher, A
Fossati, CA
机构
[1] Univ Buenos Aires, Fac Farm & Bioquim, Inst Estudios Inmunidad Humoral IDEHU, RA-1113 Buenos Aires, DF, Argentina
[2] Tech Univ Munich, Dept Chem, D-85747 Garching, Germany
关键词
D O I
10.1099/00222615-48-9-833
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Previous studies have shown that the detection of antibodies to an 18-kDa cytoplasmic protein of Brucella spp. is useful for the diagnosis of human and animal brucellosis, This protein has now been expressed in recombinant form in Escherichia coli. The recombinant protein is soluble only under reducing conditions, but alkylation with iodoacetamide renders it soluble in non-reducing media. As shown by gel exclusion chromatography, this soluble form arranges in pentamers of 90 kDa. The reactivity of human and animal sera against the recombinant protein was similar to that found with the native protein present in brucella cytoplasmic fraction, suggesting that the recombinant protein is correctly folded. The protein has low but significant homology (30%) with lumazine synthases involved in bacterial riboflavin biosynthesis, which also arrange as pentamers. Biological tests on the crude extract of the recombinant bacteria and on the purified recombinant protein showed that the biological activity of the Brucella spp, 18-kDa protein is that of lumazine synthase, Preliminary crystallographic analysis showed that the Brucella spp. lumazine synthase arranges in icosahedric capsids similar to those formed by the lumazine synthases of other bacteria. The high immunogenicity of this protein, potentially useful for the design of acellular vaccines, could be explained by this polymeric arrangement.
引用
收藏
页码:833 / 839
页数:7
相关论文
共 21 条
[1]   Diagnosis of canine brucellosis by detection of serum antibodies against an 18 kDa cytoplasmic protein of Brucella spp. [J].
Baldi, PC ;
Wanke, MM ;
Loza, ME ;
Monachesi, N ;
Fossati, CA .
VETERINARY MICROBIOLOGY, 1997, 57 (2-3) :273-281
[2]   Humoral immune response against lipopolysaccharide and cytoplasmic proteins of Brucella abortus in cattle vaccinated with B-abortus S19 or experimentally infected with Yersinia enterocolitica serotype 0:9 [J].
Baldi, PC ;
Gimbartolomei, GH ;
Goldbaum, FA ;
Abdon, LF ;
Velikovsky, CA ;
Kittelberger, R ;
Fossati, CA .
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 1996, 3 (04) :472-476
[3]   CLONING, EXPRESSION, AND OCCURRENCE OF THE BRUCELLA CU-ZN SUPEROXIDE-DISMUTASE [J].
BRICKER, BJ ;
TABATABAI, LB ;
JUDGE, BA ;
DEYOE, BL ;
MAYFIELD, JE .
INFECTION AND IMMUNITY, 1990, 58 (09) :2935-2939
[4]   STRUCTURAL CHARACTERIZATION OF THE ACTIVE-SITE OF BRUCELLA-ABORTUS CU-ZN SUPEROXIDE-DISMUTASE - A N-15 AND H-1-NMR INVESTIGATION [J].
CHEN, YL ;
PARK, S ;
THORNBURG, RW ;
TABATABAI, LB ;
KINTANAR, A .
BIOCHEMISTRY, 1995, 34 (38) :12265-12275
[5]  
DeMot R, 1996, CURR MICROBIOL, V33, P26, DOI 10.1007/s002849900068
[6]   CLONING AND SEQUENCING OF THE BACTERIOFERRITIN GENE OF BRUCELLA-MELITENSIS 16M STRAIN [J].
DENOEL, PA ;
ZYGMUNT, MS ;
WEYNANTS, V ;
TIBOR, A ;
LICHTFOUSE, B ;
BRIFFEUIL, P ;
LIMET, JN ;
LETESSON, JJ .
FEBS LETTERS, 1995, 361 (2-3) :238-242
[7]   CLONING AND NUCLEOTIDE-SEQUENCE OF THE GENE CODING FOR THE MAJOR 25-KILODALTON OUTER-MEMBRANE PROTEIN OF BRUCELLA-ABORTUS [J].
DEWERGIFOSSE, P ;
LINTERMANS, P ;
LIMET, JN ;
CLOECKAERT, A .
JOURNAL OF BACTERIOLOGY, 1995, 177 (07) :1911-1914
[8]   Protein particle vaccines against malaria [J].
Gilbert, SC ;
Hill, AVS .
PARASITOLOGY TODAY, 1997, 13 (08) :302-306
[9]   DIFFERENTIATION BETWEEN ACTIVE AND INACTIVE HUMAN BRUCELLOSIS BY MEASURING ANTIPROTEIN HUMORAL IMMUNE-RESPONSES [J].
GOLDBAUM, FA ;
RUBBI, CP ;
WALLACH, JC ;
MIGUEL, SE ;
BALDI, PC ;
FOSSATI, CA .
JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (03) :604-607
[10]   CHARACTERIZATION OF AN 18-KILODALTON BRUCELLA CYTOPLASMIC PROTEIN WHICH APPEARS TO BE A SEROLOGICAL MARKER OF ACTIVE INFECTION OF BOTH HUMAN AND BOVINE BRUCELLOSIS [J].
GOLDBAUM, FA ;
LEONI, J ;
WALLACH, JC ;
FOSSATI, CA .
JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (08) :2141-2145