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151.
L. Zavalova S. Lukyanov I. Baskova E. Snezhkov S. Akopov S. Berezhnoy E. Bogdanova E. Barsova E. D. Sverdlov 《Molecular & general genetics : MGG》1996,253(1-2):20-25
We previously detected in salivary gland secretions of the medicinal leech (Hirudo medicinalis) a novel enzymatic activity, endo-ɛ(γ-Glu)-Lys isopeptidase, which cleaves isopeptide bonds formed by transglutaminase (Factor
XIIIa) between glutamine γ-carboxamide and the ɛ-amino group of lysine. Such isopeptide bonds, either within or between protein
polypeptide chains are formed in many biological processes. However, before we started our work no enzymes were known to be
capable of specifically splitting isopeptide bonds in proteins. The isopeptidase activity we detected was specific for isopeptide
bonds. The enzyme was termed destabilase. Here we report the first purification of destabilase, part of its amino acid sequence,
isolation and sequencing of two related cDNAs derived from the gene family that encodes destabilase proteins, and the detection
of isopeptidase activity encoded by one of these cDNAs cloned in a baculovirus expression vector. The deduced mature protein
products of these cDNAs contain 115 and 116 amino acid residues, including 14 highly conserved Cys residues, and are formed
from precursors containing specific leader peptides. No homologous sequences were found in public databases.
Received: 9 April 1996 / Accepted: 17 May 1996 相似文献
152.
Development of microsatellite markers and characterization of simple sequence length polymorphism (SSLP) in rice (Oryza sativa L.) 总被引:26,自引:0,他引:26
Microsatellite markers containing simple sequence repeats (SSR) are a valuable tool for genetic analysis. Our objective is to augment the existing RFLP map of rice with simple sequence length polymorphisms (SSLP). In this study, we describe 20 new microsatellite markers that have been assigned to positions along the rice chromosomes, characterized for their allelic diversity in cultivated and wild rice, and tested for amplification in distantly related species. Our results indicate that the genomic distribution of microsatellites in rice appears to be random, with no obvious bias for, or clustering in particular regions, that mapping results are identical in intersubspecific and interspecific populations, and that amplification in wild relatives ofOryza sativa is reliable in species most closely related to cultivated rice but becomes less successful as the genetic distance increases. Sequence analysis of SSLP alleles in three relatedindica varieties demonstrated the clustering of complex arrays of SSR motifs in a single 300-bp region with independent variation in each. Two microsatellite markers amplified multiple loci that were mapped onto independent rice chromosomes, suggesting the presence of duplicated regions within the rice genome. The availability of increasing numbers of mapped SSLP markers can be expected to increase the power and resolution of genome analysis in rice. 相似文献
153.
Genomic complexity and plasticity of Burkholderia cepacia 总被引:6,自引:1,他引:5
Thomas G. Lessie William Hendrickson Brendan D. Manning Richard Devereux 《FEMS microbiology letters》1996,144(2-3):117-128
Abstract Burkholderia cepacia has attracted attention because of its extraordinary degradative abilities and its potential as a pathogen for plants and for humans. This bacterium was formerly considered to belong to the genus Pseudomonas in the γ-subclass of the Proteobacteria , but recently has been assigned to the β-subclass based on rrn gene sequence analyses and other key phenotypic characteristics. The B. cepacia genome is comprised of multiple chromosomes and is rich in insertion sequences. These two features may have played a key role in the evolution of novel degradative functions and the unusual adaptability of this bacterium. 相似文献
154.
Highly repetitive sequences and characteristics of genomic DNA in unicellular cyanobacterial strains 总被引:4,自引:0,他引:4
Munehiko Asayama Mamiko Kabasawa Isamu Takahashi Tokujiro Aida Makoto Shirai 《FEMS microbiology letters》1996,137(2-3):175-181
Abstract Microcystis aeruginosa (Synechocystis ) is a unicellular cyanobacterium that performs oxygenic photosynthesis. We found two novel sets of repetitive sequences, A (REP-A) and B (REP-B), on the M. aeruginosa K-81 genomic DNA, which consisted of distinct motifs of tandem repeated sequences located in the up- and downstream regions of the orf1 structural gene, respectively. Genomic Southern hybridization revealed multicopies of REP-A and -B on the genome. Furthermore, genomic Southern blots of cyanobacteria species with the REP-A and -B probes revealed that different hybridization signals appeared on the genomic DNAs of all 12 Microcystis strains, but no signal appeared on those of Synechocystis sp. PCC 6803, Synechococcus sp. PCC 7942, and Anabaena sp. PCC 7120. 相似文献
155.
The size relationship between the crown area of the lower canines (Cs), third (P3s) and fourth premolars (P4s), and first
molars (M1s) in hominids is examined by means of the regression analysis. The lower P3 seem to be under the influence of those
factors that control both the size of the anterior and posterior teeth, and the P4:P3 size ratio is related to the relative
size of the anterior and posterior dentitions. So, the P4>P3 sequence is associated with the megadontia and hipermegadontia
of the posterior teeth, whereas the expansion of the anterior teeth produces the P3>P4 sequence. We consider the P4:P3 size
ratio as an excellent indicator of the taxonomic and philogenetic status of fossil hominids. 相似文献
156.
Hiroyuki Yamaguchi Hitoshi Miura Kazuoki Ohsumi Takako Osaki Haruhiko Taguchi Tomoko Yamamoto Tomoko Hanawa Sachio Ogata Shigeru Kamiya 《Microbiology and immunology》1996,40(1):77-80
To determine amino acid sequences of the epitopes recognized by monoclonal antibodies (mAbs) 3C8 and 5C3 directed against Yersinia enterocolitica heat-shock protein (HSP60), a dot blot analysis was perfomed using synthesized peptides of Y. enterocolitica HSP60 such as peptides p316-342, p327-359, p340-366, p316-326, p316-321, p319-323, and p321-326 which represent positions of amino acids in Y. enterocolitica HSP60. The dot blot analysis revealed that 5C3 mAb reacted with p316-342, p316-326 and p321-326, and 3C8 mAb p316-342 and p316-326. These results indicate that the epitopes recognized by the mAbs were associated with eleven amino acids, Asp Leu Gly Gln Ala Lys Arg Val Val Ile Asn, of p316-326. The sequence homology between p316-326 of Y. enterocolitica HSP60 and the rest of the HSP60 family suggests that the five amino acids of Lys, Arg, Val, Ile and Asn, which are highly conserved in the HSP60 family, might be related with the epitope recognized by 3C8. In contrast, it was also demonstrated that three amino acids of Leu, Gly and Val, which are not well conserved in the HSP60 family, might be related to the epitope recognized by 5C3. 相似文献
157.
158.
Umesh C. Haldar Sanat K. Saha Ronald C. Beavis Nirmal K. Sinha 《Journal of Protein Chemistry》1996,15(2):177-184
Two trypsin inhibitors, LA-1 and LA-2, have been isolated from ridged gourd (Luffa acutangula Linn.) seeds and purified to homogeneity by gel filtration followed by ion-exchange chromatography. The isoelectric point is atpH 4.55 for LA-1 and atpH 5.85 for LA-2. The Stokes radius of each inhibitor is 11.4 å. The fluorescence emission spectrum of each inhibitor is similar to that of the free tyrosine. The biomolecular rate constant of acrylamide quenching is 1.0×109 M–1 sec–1 for LA-1 and 0.8 × 109 M–1 sec–1 for LA-2 and that of K2HPO4 quenching is 1.6×1011 M–1 sec–1 for LA-1 and 1.2×1011M–1 sec–1 for LA-2. Analysis of the circular dichroic spectra yields 40%-helix and 60%-turn for La-1 and 45%-helix and 55%-turn for LA-2. Inhibitors LA-1 and LA-2 consist of 28 and 29 amino acid residues, respectively. They lack threonine, alanine, valine, and tryptophan. Both inhibitors strongly inhibit trypsin by forming enzymeinhibitor complexes at a molar ratio of unity. A chemical modification study suggests the involvement of arginine of LA-1 and lysine of LA-2 in their reactive sites. The inhibitors are very similar in their amino acid sequences, and show sequence homology with other squash family inhibitors. 相似文献
159.
Computational sequence analysis of matrix metalloproteinases 总被引:12,自引:0,他引:12
Matrix metalloproteinases (MMP) play a cardinal role in the breakdown of extracellular matrix involved in a variety of biological and pathological processes. Research on MMPs has classified and characterized these enzymes according to their matrix substrate specificity, gene and protein domain structure, and regulation of activity and expression. However, the discovery of new MMPs has introduced a need for a more comprehensive and systematic method of classification and quantitative comparison of known and newly discovered members. This study compiles a sequence alignment, constructs a dendrogram, and calculates physical data and homology percentage assignments in order to obtain further insight into MMP structure-function relationships. Thorough analysis of MMP primary sequence domains, physical data patterns, and statistical analysis of sequence homology yields higher resolution in the similarities and differences that group MMP members. 相似文献
160.
Barbara Kroczynska Rengang Zhou Clifford Wood Jan A. Miernyk 《Plant molecular biology》1996,31(3):619-629
The nucleotide sequence of a cDNA clone fromArabidopsis thaliana ecotype Columbia was determined, and the corresponding amino sequence deduced. The open reading frame encodes a protein, AtJ1, of 368 residues with a molecular mass of 41 471 Da and an isoelectric point of 9.2. The predicted sequence contains regions homologous to the J- and cysteine-rich domains ofEscherichia coli DnaJ, but the glycine/phenylalanine-rich region is not present. Based upon Southern analysis,Arabidopsis appears to have a singleatJ1 structural gene. A single species of mRNA, of 1.5 kb, was detected whenArabidopsis poly(A)+ RNA was hybridized with theatJ1 cDNA. The function ofatJ1 was tested by complementation of adnaJ deletion mutant ofE. coli, allowing growth in minimal medium at 44°C. The AtJ1 protein was expressed inE. coli as a fusion with the maltose binding protein. This fusion protein was purified by amylose affinity chromatography, then cleaved by digestion with the activated factor X protease. The recombinant AtJ1 protein was purified to electrophoretic homogeneity.In vitro, recombinant AtJ1 stimulated the ATPase activity of bothE. coli DnaK and maize endosperm cytoplasmic Stress70. The deduced amino acid sequence of AtJ1 contains a potential mitochondrial targeting sequence at the N-terminus. Radioactive recombinant AtJ1 was synthesized inE. coli and purified. When the labeled protein was incubated with intact pea cotyledon mitochondria, it was imported and proteolytically processed in a reaction that depended upon an energized mitochondrial membrane.Abbreviations MBP
maltose binding protein
- PCR
polymerase chain reaction
- Stress70c
the cytosolic member of the 70 kDA family of stress-related proteins 相似文献