The DNA sequence of the structural gene of gonococcal protein III and the flanking region containing a repetitive sequence. Homology of protein III with enterobacterial OmpA proteins.

Gotschlich, Emil C.; Seiff, Michael E.; Blake, M. S.

doi:10.1084/jem.165.2.471

Cited by 70 publications

(35 citation statements)

References 50 publications

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“…However, the cysteinyl residues of PIII would be primarily in the oxidized state; they would not be labeled with mB but would be subsequently reduced by ME. These observations are consistent with the previously published characterization of PIII (8,20).…”

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confidence: 94%

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

et al. 1993

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By using a method that labels sulfhydryl-containing proteins in situ, we (6,14,15). It reacts selectively with the thiol anion of sulfhydryls, crosses biological membranes, and does not significantly alter the molecular weight of proteins (6,14,15).We have used mB to label the sulfhydryl-containing proteins of the outer membrane of Neissena gonorrhoeae. By labeling during growth, we have eliminated the effect of oxidative changes that occur during harvesting and preparation of fractions, thereby detecting the sulfhydryl status of proteins in situ.A nonpiliated, opaque colonial variant of N. gonorrhoeae 1384 (22, 23) was used for all studies. Growth conditions were as previously described (23) (Pharmacia, Piscataway, N.J.) with 0.9 mM dithiothreitol for 30 min and 5.4 mM mB for 20 min. The treated standards were diluted into sample buffer with mercaptoethanol (ME).Electrophoretic analysis of outer membranes from labeled samples revealed multiple fluorescent bands, with the two major fluorescent bands having apparent molecular masses of 41 and 31.5 kDa (Fig. 1A and B). On the basis of its intense fluorescence, the protein at 41 kDa is the major sulfhydryl-containing protein in the outer membrane. Gels stained for protein also showed the 41-kDa protein to be a major protein band in labeled fractions (Fig. 1D, lanes 3 and 5), with an intensity equivalent to that of PIII as seen in outer membrane fractions prepared by conventional methods. However, unlabeled fractions that were not reduced by ME showed no evidence of this protein (Fig. 1D, lanes 1 and 4). When unlabeled samples were reduced, only a lightly staining protein band at 41 kDa was seen (Fig. 1C, lanes 1 and 4). Samples obtained from different experiments (Fig. 1, lanes 1 to 3 versus lanes 4 and 5) confirmed the differences in the 41-kDa protein between labeled and unlabeled samples. Although there was some variation in the intensity of the 41-kDa protein between the two unlabeled, reduced samples, in both cases there was significantly less protein than in labeled samples.There are several possible reasons for the difference between the labeled and unlabeled samples. A likely explanation, given the nature of sulfhydryl compounds, the nature of the reaction with mB, and the effect of ME on the 41-kDa protein in the unlabeled samples, is that in the unlabeled samples artifacts of distribution occur during preparation of the fractions and result in decreases of the 41-kDa protein in the outer membrane; i.e., the protein undergoes autoxidation, with a change in conformation and loss from the membrane, whereas in labeled samples the 41-kDa protein is stable because of its reaction with mB. Another possibility is

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confidence: 94%

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

et al. 1993

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“…The OmpA homolog of Actinobacillus actinomycetemcomitans, Omp34, is an Fc-␥ receptor and may serve to interfere with immunoglobulin-mediated complement fixation or opsonization (19). The PIII protein of Neisseria gonorrhoeae has sequence homology with the E. coli OmpA protein (12). PIII binds blocking antibodies that interfere with the bactericidal activity of antibodies directed to other surface antigens (25).…”

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confidence: 99%

Evaluation of an Isogenic Major Outer Membrane Protein-Deficient Mutant in the Human Model of Haemophilus ducreyi Infection

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Al‐Tawfiq²,

Fortney³

et al. 2000

Infect Immun

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“…For pilin genes, a large number of stored partial gene copies contain information for variant pilin species; changes in the expressed pilin protein occur when different stored information is recombined into an expression locus (15,35,48). Repeated sequence elements located outside coding sequences have also been described, although their function is not known (7,9,10,14,15,34). With no genetic map, it has been impossible to ask questions about the organization * Corresponding author.…”

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Physical map of the chromosome of Neisseria gonorrhoeae FA1090 with locations of genetic markers, including opa and pil genes

et al. 1991

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A physical map of the chromosome of Neisseria gonorrhoeae FA1090 has been constructed. Digestion of strain FA1090 DNA with NheI, SpeI, BgM, or Pacl resulted in a limited number of fragments that were resolved by contour-clamped homogeneous electric field electrophoresis. The estimated genome size was 2,219 kb. To construct the map, probes corresponding to single-copy chromosomal sequences were used in Southern blots of digested DNA separated on pulsed-field gels, to determine how the fragments from different digests overlapped. Some of the probes represented identified gonococcal genes, whereas others were anonymous cloned fragments of strain FA1090 DNA. By using this approach, a macrorestriction map of the strain FA1090 chromosome was assembled, and the locations of various genetic markers on the map were determined. Once the map was completed, the repeated gene families encoding Opa and pilin proteins were mapped. The 11 opa loci of strain FA1090 were distributed over approximately 60% of the chromosome. The pil loci were more clustered and were located in two regions separated by approximately one-fourth of the chromosome.

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The DNA sequence of the structural gene of gonococcal protein III and the flanking region containing a repetitive sequence. Homology of protein III with enterobacterial OmpA proteins.

Cited by 70 publications

References 50 publications

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

A dominant sulfhydryl-containing protein in the outer membrane of Neisseria gonorrhoeae

Evaluation of an Isogenic Major Outer Membrane Protein-Deficient Mutant in the Human Model of Haemophilus ducreyi Infection

Physical map of the chromosome of Neisseria gonorrhoeae FA1090 with locations of genetic markers, including opa and pil genes

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