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M Vaara

Publications and source records attributed to M Vaara.

At least 73 records · Page 4Linked to original sources
PubMed1992

Polymyxin B octapeptide and polymyxin B heptapeptide are potent outer membrane permeability-increasing agents.

Polymyxin B octapeptide (PBOP) and polymyxin B heptapeptide (PBHP) were found to be effective permeabilizers of the outer membrane of Escherichia coli and Salmonella typhimurium. PBOP was as effective as polymyxin B nonapeptide (PMBN), the known very potent permeabilizer. As low a PBOP concentration as 1 microgram/ml sensitized E. coli to rifampicin by a factor of 100. Three micrograms of PBOP per ml was sufficient to sensitize this target to all the other tested hydrophobic antibiotics (erythromycin, fusidic acid, clindamycin, and novobiocin) by a factor of 30. Only a slightly higher (3-fold) concentration of PBHP was required for a similar sensitizing effect.

Amino Acid Sequence↗
PubMed1991

The nucleotide and deduced amino acid sequence of the cationic 19 kDa outer membrane protein OmpH of Yersinia pseudotuberculosis.

The OmpH proteins of enteric bacteria are recently described, small (16 kDa), cationic outer membrane proteins. Because a Yersinia pseudotuberculosis cell envelope protein of this size has been found to cross-react serologically with the human histocompatibility antigen HLA-B27 (B*2701), the sequence of Y. pseudotuberculosis OmpH was determined by sequencing the gene region which encodes mature OmpH. A protein consisting of 143 amino acid residues was found. It was 96% homologous with the OmpH of Y. enterocolitica and 62% homologous with that of Escherichia coli. Two separate OmpH regions had sequence similarity with B*2701; they were identical in both Yersinia species.

Amino Acid Sequence↗
PubMed1991

The Ssc protein of enteric bacteria has significant homology to the acyltransferase Lpxa of lipid A biosynthesis, and to three acetyltransferases.

The Ssc protein, a novel essential protein affecting the function of the enterobacterial outer membrane, matched in a protein homology search best with LpxA (UDP-N-acetylglucosamine 3-hydroxymyristoyl transferase), the enzyme which catalyzes the first step of lipid A biosynthesis. The corresponding genes, located 0.56 kb apart, were 46.7% identical. The search also revealed homology to the bacterial acetyltransferases LacA and NodL, as well as to a hypothetical protein Yglm. The region of residues 109-149 Ssc displayed the highest homology and was also homologous with another bacterial acetyltransferase, CysE, and three other bacterial proteins, two of which are hypothetical. This region and the corresponding regions of all other proteins were found to have a peculiar repeated hexapeptide pattern. Each hexapeptide unit starts with isoleucine (or its equivalent leucine and valine). In most units, the second residue is glycine and the fifth residue either valine or alanine.

Acetyltransferases↗
PubMed1991

Identification and sequence analysis of the gene mutated in the conditionally lethal outer membrane permeability mutant SS-C of Salmonella typhimurium.

The biosynthesis and structure-function relationships of the enterobacterial outer membrane are subjects of current intensive research. We have previously described the antibiotic supersensitive SS-C mutant (SH7622) of Salmonella typhimurium and shown that its outer membrane permeability barrier against hydrophobic antibiotics is severely defective. In this study, we show that this mutant is heat-sensitive, conditionally lethal, and carries a missense base-pair substitution in a novel gene which we have recently reported and now named the ssc gene. ssc encodes an earlier uncharacterized 36 kd protein (the Ssc protein) and the mutant expresses Ssc which has valine 291 changed to methionine in a methionine-rich region of Ssc. A plasmid containing the wild-type ssc allele completely reverts the antibiotic- and heat-sensitive phenotype of the SS-C mutant. Corresponding plasmids carrying the mutant allele, or an identical mutant allele prepared by localized mutagenesis, are inactive. The ssc gene is probably analogous to the firA locus of Escherichia coli which has earlier been implicated in a totally different function, mRNA synthesis. Furthermore, ssc apparently lies very close to the lpx genes involved in the thus far known steps of lipid A biosynthesis (the distance, approximately 560 bp). To conclude, our findings define a new essential gene involved in the generation of the outer membrane.

Amino Acid Sequence↗
PubMed1991

Molecular mimickry between HLA B27 and Yersinia, Salmonella, Shigella and Klebsiella within the same region of HLA alpha 1-helix.

Two new examples of amino acid homology between HLA B27 and microbes triggering HLA B27-associated diseases are described. An outer membrane protein YadA (Yersinia adhesin, previously called Yop1) of Yersinia enterocolitica and Y. pseudotuberculosis shares a linear tetrapeptide with HLA B27. A cationic outer membrane protein OmpH of Salmonella typhimurium shares homology with five amino acids of HLA B27 in a non-linear fashion. The four amino acids of YadA are also notably included in the hexapeptide identical between Klebsiella pneumoniae nitrogenase and HLA B27, and three of them occur in the pentapeptide shared by a Shigella flexneri protein and HLA B27. Antibodies against synthetic peptides including HLA B27 homologues sequences of YadA and OmpH were observed in one-third of the patients with HLA B27 associated diseases. Antibodies were directed against a flanking sequence next to the amino acid sequences shared by arthritis-triggering microbes and HLA B27. The area of identity in each example of this molecular mimicry (Yersinia, Salmonella, Shigella and Klebsiella) is located in the same place on the HLA B27 molecule: between amino acids 70 to 78 in the variable region of alpha 1-helix. This area of HLA B27 molecule includes sites predicted to be important for binding processed antigens.

Adhesins, Bacterial↗
PubMed1991

Polyamines as constituents of the outer membranes of Escherichia coli and Salmonella typhimurium.

Extraction of whole cells of Salmonella typhimurium and Escherichia coli with 1 M NaCl released 8 to 13% of their total cellular polyamines (putrescine, cadaverine, and spermidine). This extraction did not cause significant cell lysis, release of outer membrane (OM) constituents, or leakage of periplasmic beta-lactamase. The extraction released nearly equal amounts of polyamines from mdo (membrane-derived oligosaccharide) mutants and wild type. These findings suggest that the released polyamines are apparently bound to the cell envelope. NaCl (1 M) was as effective as trichloroacetic acid in releasing polyamines from isolated OM and lipopolysaccharide (LPS). Isolated OM contained four times more polyamines than the cytoplasmic membrane. The increased binding to the OM is apparently due to the association of polyamines with the polyanionic LPS. Nearly identical amounts of polyamines were found in the OM and LPS preparations (as quantified per milligram of LPS). These amounts are equal to those released from the intact cells by 1 M NaCl (quantitation as above). However, redistribution of polyamines took place after cell disruption, because the relative proportions of different polyamines varied in the OM and LPS preparations. These results indicate that polyamines released from intact cells during 1 M NaCl extraction are preferentially derived from the OM.

Azides↗
PubMed1991

The ompH gene of Yersinia enterocolitica: cloning, sequencing, expression, and comparison with known enterobacterial ompH sequences.

We have recently described a previously uncharacterized outer membrane protein of Salmonella typhimurium and Escherichia coli and cloned and sequenced the corresponding gene, the ompH gene, of S. typhimurium (P. Koski, M. Rhen, J. Kantele, and M. Vaara, J. Biol. Chem. 264:18973-18980, 1989). We report here the cloning, sequencing, and expression of the corresponding gene of Yersinia enterocolitica. It is significantly homologous to the ompH genes of E. coli and S. typhimurium (homology percentages, 65 and 64%, respectively), has a promoter region strongly homologous to the E. coli 17-bp class consensus promoter, and encodes a protein consisting of 165 amino acids (22 of which form the signal sequence). The plasmid-borne Y. enterocolitica ompH was found to be expressed both in the E. coli host and in minicells. The isolated outer membrane of Y. enterocolitica was shown to contain OmpH. The homology of the Y. enterocolitica OmpH protein is 66% with E. coli OmpH and 64% with S. typhimurium OmpH. All OmpH proteins have almost identical hydrophobic profiles, charge distributions, and predicted secondary structures. Because yersiniae are considered rather distant relatives of E. coli and S. typhimurium in the Enterobacteriaceae family, these results might indicate that most or all strains of the family Enterobacteriaceae have OmpH proteins remarkably homologous to those now sequenced.

Amino Acid Sequence↗
PubMed1991

The outer membrane permeability-increasing action of deacylpolymyxins.

The outer membrane permeability-increasing action of deacylpolymyxins was compared to the well-known potent action of polymyxin B nonapeptide (PMBN). Deacylpolymyxin B (DAPB), prepared by treating polymyxin B with polymyxin acylase, was found to be a slightly more effective permeabilizer than PMBN. As low a DAPB concentration as 1 microgram/ml sensitized Escherichia coli to the probe antibiotics (rifampin, fusidic acid, erythromycin, clindamycin, novobiocin) by factors 30-100 and Salmonella typhimurium by factors 10-100. A higher concentration (3 micrograms/ml) of DAPB elicited further sensitization. Also deacylcolistin (DAC) was found to be an effective permeabilizer.

Amino Acid Sequence↗
PubMed1991

The outer membrane permeability-increasing action of linear analogues of polymyxin B nonapeptide.

Polymyxin nonapeptides such as polymyxin B nonapeptide (PMBN) are polymyxin-derived deacylated nonapeptides which contain a heptapeptide ring and are known as effective permeabilizers of the outer membrane (OM) of Gram-negative bacteria. In order to assess the role of the cyclic moiety of PMBN in the permeabilization of the OM, the author compared the OM permeabilizing activity of two synthetic linear PMBN analogues with the well-characterized activity of PMBN. While a low concentration (1-3 micrograms/ml) of PMBN was sufficient to sensitize both Escherichia coli and Pseudomonas aeruginosa to hydrophobic probe antibiotics (rifampin, fusidic acid) by a factor of 100, even a high concentration (100 micrograms/ml) of linear arginyl polymyxin B decapeptide sensitized E. coli only by a factor of 3 and did not sensitize P. aeruginosa at all. In identical assays, linear lysyl polymyxin B nonapeptide completely lacked any sensitizing activity. These findings indicate that the cyclic peptide ring is crucial for the OM-permeabilizing activity of polymyxin nonapeptides.

Amino Acid Sequence↗
PubMed1990

Primary structure and expression of the Ssc-protein of Salmonella typhimurium.

A 1020-bp open reading frame (ORF) was found immediately downstream of the ompH gene of Salmonella typhimurium. This ORF (ORF-36) encodes a moderately hydrophobic protein with 341 amino acid residues (calculated molecular mass, 35,928 Da). The ORF-36 product was detected in minicells. Downstream of ORF-36, another ORF was found. It is highly homologous to the E. coli ORF (ORF-17.4) which precedes the lpx-genes involved in lipid A biosynthesis. ORF-36 is probably analogous to the firA gene of E. coli, the sequence of which has not yet been published. Thus it appears that the enterobacterial ompH and lpx genes are separated only by the ORF-36 and ORF-17.4 genes. We also discuss the data on the function of the ORF-36 protein. On this basis, we suggest that the protein could be called the Ssc protein.

Acetyltransferases↗
PubMed1990

Partitioning of hydrophobic probes into lipopolysaccharide bilayers.

Lipophilic solutes permeate rapidly through lipid bilayer membranes. However, the outer membrane of enteric bacteria, which is composed of a lipopolysaccharide monolayer outer leaflet and the glycerophospholipid inner leaflet, shows extremely low permeability to hydrophobic solutes. In order to examine the cause of this exceptionally low permeability, the lipid/water partition behavior of various lipophilic probes was determined by using lipopolysaccharides of various chemotypes and glycerophospholipids. With all probes, under many different conditions, the lipopolysaccharide/water partition coefficients were generally about an order of magnitude smaller than the phospholipid/water partition coefficients, and this result is consistent with the low permeability of the lipopolysaccharide monolayer, and hence the asymmetric bilayer found in the outer membrane. Furthermore, organic polycations significantly increased the partition of N-phenylnaphthylamine into lipopolysaccharides, a result again consistent with the permeability-increasing effect of such cations on intact outer membrane. Very defective, 'deep rough' lipopolysaccharides of chemotypes Rd2, Rd1 and Re, had only slightly (20-75%) higher partition coefficients in comparison with the more complete lipopolysaccharides, and this difference is probably not enough to explain the approximately 100-fold increase in lipophile permeability seen in deep rough strains.

1-Naphthylamine↗
PubMed1990

Complete sequence of the ompH gene encoding the 16-kDa cationic outer membrane protein of Salmonella typhimurium.

The complete nucleotide sequence of the ompH gene encoding the 16-kDa basic outer membrane protein of Salmonella typhimurium was determined. The OmpH protein is synthesized in a precursor form with additional 20 amino acid residues in the N terminus of the protein. This peptide has common characteristics of signal sequences. The promoter region has strong homology to consensus sequences of Escherichia coli. The expression of ompH was detected in minicells.

Amino Acid Sequence↗
PubMed1990

Bacterial 'histone-like protein I' (HLP-I) is an outer membrane constituent?

The nucleoid-associated 'histone-like protein I' (HLP-I) protein of E. coli was found to be homologous with the cationic 16-kDa outer membrane protein OmpH of Salmonella typhimurium. Deduced from the nucleotide sequence, the HLP-I protein has 91% identical residues with the OmpH protein. Both proteins have very similar cleavable signal sequences. The nucleotide sequence similarity between the corresponding genes hlpA and ompH is 87%. The ompH gene is located in a gene cluster resembling the hlpA-ORF17 region of E. coli which is close to the Ipx genes involved in the biosynthesis of lipopolysaccharides. The localization of the OmpH/HLP-I protein in the cell is discussed.

Amino Acid Sequence↗
PubMed1990

The effect of oligolysines Lys-3, Lys-4, and Lys-5 on the outer membrane permeability of Pseudomonas aeruginosa.

A lysine polymer with five residues (Lys-5) was found to remarkably increase the outer membrane (OM) permeability of Pseudomonas aeruginosa to the tested hydrophobic probes (nitrocefin, N-phenyl naphthylamine, rifampin). Lys-3 and Lys-4 were inactive. The OM of Escherichia coli and Salmonella typhimurium was not permeabilized by Lys-5. Furthermore, even the action of Lys-5 on the Pseudomonas OM was abolished when the assays were performed in the presence of 150 mM NaCl instead of the low-ionic strength buffer earlier used by investigators studying the effect of polycations on the Pseudomonas OM.

1-Naphthylamine↗
PubMed1990

Antimicrobial susceptibility of Salmonella typhimurium carrying the outer membrane permeability mutation SS-B.

The antibiotic susceptibility profile of Salmonella typhimurium SS-B, a mutant susceptible to some antimicrobial agents, was studied in detail. Twenty-eight agents were tested, and eleven of these had MICs significantly lower (32- to greater than 250-fold) for the SS-B strain than for its parent. The drugs were generally hydrophobic or amphiphilic. Polymyxin B nonapeptide, which has a known outer membrane permeabilizing action, further reduced the MIC of several of these agents for the SS-B strain by a factor of approximately 10 to 30. In most cases, the resulting MICs were lower than the corresponding MICs for the parent strain grown in the presence of polymyxin B nonapeptide. In addition, the hydrophobic fluorescent probe N-phenyl naphthylamine was rapidly embedded in the membranes of the SS-B strain but was poorly embedded in those of the parent strain.

Anti-Bacterial Agents↗
PubMed1990

Do salicylates and ascorbate increase the outer membrane permeability to hydrophobic antibiotics in Pseudomonas aeruginosa?

Acetylsalicylate and ascorbate have earlier been shown to increase the outer membrane (OM) permeability of Pseudomonas aeruginosa to a hydrophobic probe compound, nitrocefin. In order to elucidate whether these drugs increase the OM permeability to a wider set of hydrophobic compounds, the OM permeability to three other hydrophobic probes (rifampin, fusidic acid and sodium deoxycholate) was studied in the presence of salicylates or ascorbate. A high concentration (300 micrograms/ml, equal to 1.7 mM) of L-ascorbate decreased the minimum inhibitory concentration (MIC) of rifampin against P. aeruginosa by a factor of approximately 3. As a sharp contrast, the reference compound, polymyxin B nonapeptide (PMBN) which has a strong OM permeability-increasing action, decreased the MIC by a factor of approximately 100, at a concentration as low as 3 micrograms/ml (equal to 3 microM). If the assays were performed in a low ionic strength medium (L broth diluted 1/5 with water) instead of L broth, ascorbate was somewhat more effective. The MIC of fusidic acid was even less influenced by ascorbate. Additionally, ascorbate did not potentiate the bacteriolytic action of sodium deoxycholate, whereas the control compound hexametaphosphate had a marked effect. Furthermore, salicylate and acetylsalicylate sensitised, in all conditions tested, P. aeruginosa to none of the three probes. The results suggest that ascorbate and salicylates lack any significant OM permeability-increasing action.

Ascorbic Acid↗