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A Isogai

Publications and source records attributed to A Isogai.

At least 55 records · Page 3Linked to original sources
PubMed1996

The SLGs corresponding to the same S24-haplotype are perfectly conserved in three different self-incompatible Brassica campestris L.

We have identified three strains of the same S24-haplotype in self-incompatible B. campestris L. Two of them, S-12j and 1-1j, have been derived from Japanese populations and one, 27-1t, from Turkish one. The cDNA clones of SLG24 (S24-locus glycoprotein), which linked to the S-locus, were isolated from each strain, and sequenced. Each clone isolated from S-12j, 1-1j and 27-1t, was designated as SLG24 (S-12j), SLG24 (1-1j) and SLG24 (27-1t), respectively. Their nucleotide sequences were completely identical in coding region, 5' non-coding region, and 3' non-coding region, though the position of the polyadenylation site was slightly different among the cDNA clones. This result suggests that the origin of S24-haplotype in these three strains might be common, and that the nucleotide sequences of SLG24 of the same S24-haplotype are completely conserved among different populations. The high conservation of the SLG24 nucleotide sequences is probably essential for the recognition of self or non-self of self-incompatibility in this S24-haplotype.

Amino Acid Sequence↗
PubMed1995

Stereochemistry of chitin hydrolysis by a plant chitinase/lysozyme and X-ray structure of a complex with allosamidin: evidence for substrate assisted catalysis.

The plant enzyme hevamine has both chitinase and lysozyme activity. HPLC analysis of the products of the hydrolysis of chitopentaose shows that hevamine acts with retention of the configuration, despite the absence of a nucleophilic or stabilizing carboxylate. To analyze the stabilization of a putative oxocarbonium ion intermediate, the X-ray structure of hevamine complexed with the inhibitor allosamidin was determined at 1.85 A resolution. This structure supports the role of Glu127 as a proton donor. The allosamizoline group binds in the center of the active site, mimicking a reaction intermediate in which a positive charge at C1 is stabilized intramolecularly by the carbonyl oxygen of the N-acetyl group at C2.

Acetylglucosamine↗
PubMed1995

Three-dimensional solution structure of bombyxin-II an insulin-like peptide of the silkmoth Bombyx mori: structural comparison with insulin and relaxin.

The three-dimensional solution structure of bombyxin-II, an insulin-like two-chain peptide produced by the brain of the silkworm Bombyx mori, has been determined by simulated annealing calculations based on 535 distance constraints and 24 torsion-angle constraints derived from NMR data and three distance constraints of the disulfide bonds. To our knowledge, this is the first three-dimensional structure determined for an invertebrate insulin-related peptide. The root-mean-square deviations between the best 10 structures and the mean structure are 0.58(+/- 0.15) A for the backbone heavy atoms (N, C alpha, C) and 1.03(+/- 0.18) A for all non-hydrogen atom if less well-defined N and C termini (A1, A20, B(-2) to B4 and B23 to B25) are excluded. The overall main-chain structure of bombyxin-II is similar to that of insulin. However, there are significant conformational and functional differences in their B-chain C-terminal parts. The B-chain C-terminal part of bombyxin-II adopts an extension of the B-chain central helix like that of relaxin and is not required for bombyxin activity, while the corresponding part of insulin adopts a sharp turn and a beta-strand and is essential for insulin activity. This structure demonstrates that bombyxin-II is more closely related to relaxin than to insulin, and suggests that insulin might have evolved the additional receptor-recognition site in the B-chain C-terminal beta-strand to distinguish itself from bombyxin and relaxin. The structure of bombyxin-II thus provides novel insights into the receptor recognition and divergent molecular evolution of insulin-superfamily peptides.

Amino Acid Sequence↗
PubMed1995

Identification of the receptor-recognition surface of bombyxin-II, an insulin-like peptide of the silkmoth Bombyx mori: critical importance of the B-chain central part.

Bombyxin-II, a brain-secretory peptide of the silkmoth Bombyx mori, shares 40% sequence identify and the characteristics core structure with human insulin. In spite of the structural similarity, no cross-activity is observed between them. To localize the active region of bombyxin-II, we have synthesized chimeric molecules of bombyxin-II and human insulin, and examined their bombyxin activity. Two chimeric molecules, which were sequentially identical except for the B-chain central part, showed significantly different potencies in bombyxin activity. Solution structure determination of these chimeric molecules revealed that their B-chain central parts took similar main-chain conformation, but formed dissimilar patches on their molecular surfaces. Therefore, the surface patch formed by the central part of the bombyxin-II B-chain is of critical importance for recognition of the bombyxin receptor. The above results, together with other data on the structure-activity relationships of bombyxin, indicate that the receptor-recognition surface of bombyxin-II includes the A-chain N and C, termini in addition to the B-chain central part. Though bombyxin-II, human insulin and human relaxin 2 use the common surface as their receptor-recognition sites, each of the surface patches is characterized by the variety of involved side-chains. Insulin and relaxin involve additional parts for receptor recognition, particularly the B-chain C-terminal part and the extended A-chain N-terminal helix, respectively. In conclusion, these ligands have evolved their own specific mechanisms for receptor recognition while retaining the major recognition surface.

Amino Acid Sequence↗
PubMed1995

Saprophytic Pseudomonas syringae strain M1 of wheat produces cyclic lipodepsipeptides.

A saprophytic fluorescent bacterium (strain M1) isolated from wheat was identified as Pseudomonas syringae and shown to produce the cyclic lipodepsipeptides, syringomycin E and syringopeptin SP25A. M1 grew in planta but did not affect germination or cause disease symptoms in wheat. The findings show that the production of these metabolites, generally regarded as plant virulence factors, does not correlate with plant pathogenicity.

Amino Acid Sequence↗
PubMed1995

Evolutionary aspects of the S-related genes of the Brassica self-incompatibility system: synonymous and nonsynonymous base substitutions.

In the Brassicaceae, self-vs. nonself-recognition in self-incompatibility is controlled by sporophytic S-alleles. Haplotypes specifying both SRK (S-receptor kinase) and SLG (S-locus glycoprotein) are considered to play an important role in the recognition reactions. We compared the nucleotide sequences of SRK9(Bc) and SRK6(Bo). The number of nonsynonymous substitutions per site (Pn) was lower, constrained, in the kinase than the receptor domain, while the numbers of synonymous substitutions (Ps) in the two domains were largely comparable. Pairwise values for Ps and Pn were calculated among 17 operational taxonomic units, including eight SLGs, the receptor domains of two SRKs, four SRAs (S-related A) and three SRBs (S-related B), which have high homologies with each other. The values of Ps and Pn of SLG were mostly comparable to those of the receptor domain of SRK. Dendrograms constructed on the basis of Pn and Ps indicated that SRA differentiated first, followed by SRB. The differentiation of SLG alleles is one of prerequisite factors for the establishment of self-incompatibility, and the allelic differentiation has occurred more than tens of million years ago.

Alleles↗
PubMed1995

Cloning and characterization of a region of Enterococcus faecalis plasmid pPD1 encoding pheromone inhibitor (ipd), pheromone sensitivity (traC), and pheromone shutdown (traB) genes.

Bacteriocin plasmid pPD1 in Enterococcus faecalis encodes a mating response to recipient-produced sex pheromone cPD1. Once a recipient acquires pPD1, transconjugants apparently shut off cPD1 activity in broth culture and no longer behave as recipients for pPD1. This event is performed by synthesis of the pheromone inhibitor iPD1 and also by repression of cPD1 production, the so-called "pheromone shutdown." A 5.4-kb EcoRV-HincII segment of pPD1, which expressed iPD1 in Escherichia coli, was sequenced and found to be organized as traC-traB-traA-ipd; each open reading frame is analogous to that found in other pheromone plasmids, pAD1 and pCF10, and thus is designated in accordance with the nomenclature in pAD1. The ipd gene encodes a peptide consisting of 21 amino acids, in which the C-terminal eight residues correspond to iPD1. The putative TraC product has a strong similarity to oligopeptide-binding proteins found in other bacterial species, as do pheromone-binding proteins of pCF10 and pAD1. A strain carrying traC-disrupted pPD1 required a concentration of cPD1 fourfold higher than that needed by the wild-type strain for induction of sexual aggregation. These results suggest that the TraC product contributes to pheromone sensitivity as a pheromone-binding protein. A strain transformed with traB-disrupted pPD1 produced a high level of cPD1 similar to that produced by plasmid-free recipients and underwent self-induction. Thus, the TraB product contributes to cPD1 shutdown.

Amino Acid Sequence↗
PubMed1995

Structural analysis of new syringopeptins by tandem mass spectrometry.

New syringopeptins SP(SC)-1 and -2 were isolated from culture filtrates of phytopathogenic bacterium strain SC1 of Pseudomonas syringae pv. syringae. These syringopeptins were composed of a beta-hydroxy fatty acid, a long sequence of aliphatic amino acids. and a lactone moiety of eight amino acids. The amino acid sequences were deduced from a comparison of their tandem mass sepctra with those of known syringopeptins SP-22a and SP-25a. SP(SC)-1 and SP(SC)-2 resembled SP-22a, but differed from the latter by 3 amino acids.

Amino Acid Sequence↗
PubMed1995

The sequences of S-receptor kinases (SRK) involved in self-incompatibility and their homologies to S-locus glycoproteins of Brassica campestris.

S-Receptor kinase (SRK) is a kind of receptor protein kinase and has a receptor domain resembling S-locus glycoprotein (SLG). cDNAs encoding SRK8 and SRK12 were isolated respectively from S8 and S12 haplotypes of Brassica campestris. The SLG domains of SRKs were highly homologous to SLGs, suggesting that both SLG and SRK might bind the same ligands in recognition of self-pollen.

Amino Acid Sequence↗
PubMed1995

Purification and properties of fructosyl lysine oxidase from Fusarium oxysporum S-1F4.

Fructosyl lysine oxidase (FLOD) was examined for its use in the enzymatic measurement of the level of glycated albumin in blood serum. To isolate microorganisms having such an enzyme activity, we used N epsilon-fructosyl N alpha-Z-lysine (epsilon-FL) as a sole nitrogen source in the enrichment culture medium. The isolated fungus, strain S-1F4, showed a high FLOD activity in the cell-free extract and was identified as Fusarium oxysporum. FLOD was purified to an apparent homogeneity on SDS-PAGE. The molecular mass of the subunit was 50 kDa on SDS-PAGE and seemed to exist in a monomeric form. The enzyme had an absorption spectrum characteristic of a flavoprotein and the flavin was found to be covalently bound to the enzyme. The enzyme acted against N epsilon-fructosyl N alpha-Z-lysine and N alpha-fructosyl N epsilon-Z-lysine and showed specificity for fructosyl lysine residues.

Amino Acid Oxidoreductases↗
PubMed1995

Isolation and structure of the Enterococcus faecalis sex pheromone, cOB1, that induces conjugal transfer of the hemolysin/bacteriocin plasmids, pOB1 and pYI1.

A bacterial sex pheromone, cOB1, which induces conjugal transfer of the Enterococcus faecalis hemolysin-bacteriocin (Hly/Bac) plasmid, pOB1, was isolated from the culture broth of pOB1-free E. faecalis. Its structure was found to be a hydrophobic octapeptide, H-Val-Ala-Val-Leu-Val-Leu-Gly-Ala-OH. The cOB1 peptide induced the mating response of not only pOB1 but also another incompatibility group Hly/Bac plasmid, pYI1.

Amino Acid Sequence↗
PubMed1995

Rotihibins, novel plant growth regulators from Streptomyces graminofaciens.

In the course of screening search for plant growth regulators, a culture filtrate of Streptomyces graminofaciens 3C02 was found to inhibit the growth of lettuce seedlings. The active substances, named rotihibin A (1) and B (2), were revealed to be lipo-peptidal compounds. Rotihibins inhibit growth of various plants at below 1 microgram/ml, but do not show lethal activity even at higher doses.

Molecular Structure↗
PubMed1995

Molecular cloning of members of the S-multigene family in self-incompatible Brassica campestris L.

We isolated 12 groups of genomic clones that contained SLG-homologous regions from a genomic library constructed from an S9 homozygote of self-incompatible Brassica campestris. Both SLG9 and SRK9 genomic clones, which are located within the self-incompatibility (S) locus, were included in these groups. The promoter regions of SLG9 and SRK9 were completely identical for at least 200 bp upstream from their respective initiation codons (ATG). The five sequence elements (boxes I to V) that are conserved in the promoters of SLG and SRK genes were also found in the SLG9 and SRK9 clones. However, one conserved element (box III) unexpectedly lacked 7 of 11 bp, although box III has been considered necessary for expression in pistil. The other ten groups of genomic clones were classified into six SRK-like groups and four SLG-like groups. These results indicate that SLG, SRK, SLG-like, and SRK-like genes form a large S-multigene family in B. campestris.

Base Sequence↗
PubMed1994

Assignment of disulfide bond location in prothoracicotropic hormone of the silkworm, Bombyx mori: a homodimeric peptide.

The disulfide bond location of a homodimeric peptide, prothoracicotropic hormone (PTTH) of the silkworm, Bombyx mori, was determined by a combination of partial reduction and sequence analysis of peptide fragments generated through a partial reduction of PTTH followed by alkylation and enzyme digestion. The partial reduction and S-alkylation broke the interchain disulfide bond but did not affect the intrachain disulfide bonds, generating monomeric PTTH whose intrachain disulfide bonds were kept intact. This monomeric PTTH has about one-half the biological activity of intact PTTH. Sequence analysis of the fragments generated by lysyl endopeptidase digestion of this monomeric PTTH after complete reduction and S-alkylation by another S-alkylating reagent showed that only the Cys15 residue was reduced and S-alkylated by the foregoing partial reduction, indicating that this residue formed the interchain disulfide bond. The other disulfide bonds which formed intrachain bridgings were determined by sequence and mass analyses of the fragments generated by two successive enzyme digestions of the monomeric PTTH. In conclusion, the disulfide bond location of PTTH was assigned to Cys15-Cys15' as an interchain disulfide linkage and Cys17-Cys54, Cys40-Cys96, and Cys48-Cys98 as intrachain disulfide linkages.

Amino Acid Sequence↗
PubMed1994

A high degree of homology exists between the protein encoded by SLG and the S receptor domain encoded by SRK in self-incompatible Brassica campestris L.

The sporophytic self-incompatibility system in Brassica campestris is controlled by multiple alleles of a single locus, S. We isolated and characterized cDNA clones that correspond to genes for S-glycoprotein (SLG) and S-receptor kinase (SRK) from an S9-homozygous strain. An SRK9 fragment was amplified by the polymerase chain reaction (PCR) by use of primers that corresponded to regions of the gene for the extracellular receptor domain (S-domain) and the kinase domain. The fragment was used as a probe to isolate an SRK9 cDNA clone from the cDNA library of stigmas of an S9-homozygote. An SLG9 cDNA clone was also isolated from the library by use of SLG8 cDNA as a probe. Restriction fragment length polymorphism (RFLP) linkage analysis of the F2 plants from an S8S9 hybrid demonstrated that the SRK and SLG genes were tightly linked to one another and were also linked to S-genotypes, as determined by pollination tests. The transcripts of SRK9 and SLG9 were detected in stigmas, but not in anthers or leaves. The cytoplasmic kinase domain encoded by SRK9 contained amino acid sequences that are conserved in serine/threonine kinases. The nucleotide sequence encoding the S-domain of SRK9 was 98.4% homologous to that of SLG9 at the nucleotide level. This observation suggests the existence of a mechanism for maintaining a high degree of similarity between SLG and the region that encodes the S-domain of SRK.

Base Sequence↗
PubMed1994

The prgQ gene of the Enterococcus faecalis tetracycline resistance plasmid pCF10 encodes a peptide inhibitor, iCF10.

Conjugative transfer of the Enterococcus faecalis tetracycline resistance plasmid pCF10 is stimulated by a peptide pheromone, cCF10. Once a recipient strain acquires pCF10 and thus becomes a pheromone-responsive donor, cCF10 activity is no longer detected in culture filtrates. Here we show that pCF10 encodes a peptide inhibitor, iCF10, secreted by donor cells; this inhibitor antagonizes the cCF10 activity in culture filtrates. In order to detect and quantitate iCF10, we developed a reverse-phase high-performance liquid chromatography assay in which the inhibitor peptide elutes separately from the pheromone; this type of assay enabled us to determine that lack of pheromone activity in donor culture filtrates was due to secretion of a mixture of iCF10 and cCF10, rather than abolition of cCF10 secretion. The gene encoding iCF10, prgQ, is located on the EcoRI-C fragment of pCF10. The open reading frame comprising the prgQ gene encodes a 23-amino-acid precursor that resembles a signal peptide. This precursor is cleaved to the mature heptapeptide iCF10 during the secretion process.

Amino Acid Sequence↗
PubMed1994

The sequences of S-glycoproteins involved in self-incompatibility of Brassica campestris and their distribution among Brassicaceae.

S-Glycoproteins have been thought to play a role in the self-incompatibility in Brassica plants. Two cDNAs encoding S-glycoproteins were isolated from stigmas of two strains of Brassica campestris. Their amino acid sequences are highly homologous to each other. However, the numbers of the potential glycosylation sites are different, although most of them are conserved. In the middle part of the sequences, there are two supervariable regions which might contribute to the specificity of each S-glycoprotein. Genomic Southern analysis showed many bands hybridizing with cDNA of S-glycoproteins in various Brassica species including self-compatible species.

Amino Acid Sequence↗