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M M Lieberman

Publications and source records attributed to M M Lieberman.

28 records · Page 2Linked to original sources

Pseudomonas ribosomal vaccines: preparation, properties, and immunogenicity.

The preparation, properties, and immunogenicity of ribosomal vaccines from Pseudomonas aeruginosa are described. These preparations, containing protein and RNA, were tested for immunogenicity by active immunization of mice and subsequent challenge with homologous, live bacteria. The results demonstrated that vaccines prepared from a majority of serotypes used were immunogenic, i.e., afforded 60 to 100% mouse protection against a challenge inoculum containing 8 to 50 50% lethal doses. In some cases vaccine doses as low as 1 microgram of RNA provided 100% mouse protection. Molecular sieve chromatography of a highly immunogenic ribosomal preparation on Sepharose 4B demonstrated the presence of two molecular weight fractions: (i) peak A, an excluded peak (thus having a molecular weight of at least 2 times 10(7)), and (ii) peak B, considerably retarded, with an elution position corresponding to a molecular weight of about 2.2 X 10(6), approximating that of typical 70S ribosomes. Both peaks A and B were immunogenic; however, the immunogenicity of peak A was greater (i.e., a smaller immunizing dose was required) than that of peak B. Peak A was shown to contain components of lipopolysaccharide in addition to protein and RNA (which comprised 80% of the dry weight of peak A). On the other hand, peak B was shown to be free of lipopolysaccharide, and 100% of its dry weight consisted of protein and RNA.

Animals↗

Derepression of GDP-alpha-mannose and UDP-glucose pyrophosphorylases by a regulator gene mutation; episomal dominance in partial diploids.

Mutants of Escherichia coli K12 at the capR locus are overproduce capsular polysaccharide, and are derepressed for synthesis of several enzymes involved in capsular polysaccharide synthesis(1-3) including GDP-mannose pyrophosphorylase.(8) UDP-glucose pyrophosphorylase is also derepressed in a haploid capR9 mucoid mutant. Heterozygous mucoid partial diploids with the capR9 allele on the episome and the wild-type (capR(+)) allele on the chromosome (F'capR9/capR(+)) are derepressed for UDP-glucose pyrophosphorylase and GDP-mannose pyrophosphorylase, while the reciprocal nonmucoid heterozygotes (F'capR(+)/capR9) are repressed for these enzymes. These results provide evidence that the episomal capR allele is dominant with respect to synthesis of these two enzymes.

Diploidy↗

Control of uridine diphosphate-glucose dehydrogenase synthesis and uridine diphosphate-glucuronic acid accumulation by a regulator gene mutation in Escherichia coli K-12.

Uridine diphosphate (UDP)-glucose dehydrogenase, the enzyme that converts UDP-glucose to UDP-glucuronic acid, was derepressed in a mucoid (capR9) strain of Escherichia coli K-12 and repressed in a nonmucoid (capR(+)) strain. A nonmucoid mutant (strain MC 152; capR9 non-2) derived from the mucoid strain accumulated large quantities of nucleotides. Among these nucleotides, UDP-glucuronic acid was identified as well as guanosine triphosphate and an adenosine diphosphate-sugar. UDP-glucose dehydrogenase was still derepressed in strain MC 152. When the nonmucoid mutant was transduced to the wild-type state for this regulator gene (capR(+)), the transductant was found to accumulate less total nucleotides, and the accumulation of UDP-glucuronic acid was abolished. UDP-glucose dehydrogenase was repressed in the capR(+)non-2 strain but not to the same extent that it was in the capR(+) strain.

Alcohol Oxidoreductases↗

Depression of guanosine diphosphate-mannose pyrophosphorylase by mutations in two different regulator genes involved in capsular polysaccharide synthesis in Escherichia coli K-12.

Mutations in a regulator gene (capR) that causes increased synthesis of capsular polysaccharide and derepressed synthesis of several enzymes involved in polysaccharide synthesis also derepress synthesis of guanosine diphosphate (GDP)-mannose pyrophosphorylase. In addition, a second mucoid mutation (capS, which maps separately from capR) also results in the derepression of GDP-mannose pyrophosphorylase. New conditions for assaying GDP-mannose hydrolyase and GDP-l-fucose synthetase permitted us to show that these enzymes are also derepressed in the capS mucoid strain. Although phosphomannose isomerase and uridine diphosphate-galactose-4-epimerase are derepressed in capR mucoid strains, they are not derepressed in capS mucoid strains. A nonmucoid mutant of a strain containing the capR9 (mucoid) allele was deficient in GDP-mannose pyrophosphorylase.

Cell Wall↗

Genetic and biochemical studies on mannose-negative mutants that are deficient in phosphomannose isomerase in Escherichia coli K-12.

Two mannose-negative mutants of Escherichia coli K-12 have been isolated. These mutants are deficient in the ability to synthesize phosphomannose isomerase and capsular polysaccharide when grown on glucose-containing media. Interrupted mating experiments to determine the kinetics of genetic transfer show that the two mannose-negative mutations map together between the histidine and tryptophan regions of the E. coli chromosome.

Chromosome Mapping↗

Derepression of phosphomannose isomerase by regulator gene mutations involved in capsular polysaccharide synthesis in Escherichia coli K-12.

A regulator gene mutation (capR) that causes increased synthesis of capsular polysaccharide and derepressed synthesis of several enzymes involved in polysaccharide synthesis also derepresses phosphomannose isomerase (PMI) synthesis. In contrast, a second mutation (capS, which maps separately from capR) that causes increased production of the same polysaccharide does not lead to increased synthesis of PMI (nor of several of the other enzymes involved in polysaccharide synthesis). Introduction of the capR9 allele by transduction or mutation of capR(+) to capR can change the phenotype of a mannose-negative nonmucoid strain to a mannose-positive mucoid phenotype. Thus, genotype capR(+)man-2 is mannose-negative and nonmucoid, but genotype capR9 man-2 is mannose positive and mucoid. Other interactions between these alleles in the synthesis of capsular polysaccharide are recorded.

Chromosome Mapping↗

Parallel synthesis and biocatalytic amplification of a cross-conjugated cyclopentenone library.

A combination of parallel chemical synthesis and biocatalysis has been used to prepare and amplify a library of cross-conjugated cyclopentenones. A number of marine and terrestrial natural products with antibiotic activity are known to incorporate this pharmacophore. The library was screened for anticancer, antimycobacterial, antifungal, and antibacterial activity. The positive results from the screens provide an indication of the structural features that are associated with activity in the various assays and suggest promising avenues for further inquiry.

Biotransformation↗