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171.
172.
Shechinah Felice Choragudi Ganesh Kumar Veeramachaneni BV Raman Bondili JS 《Bioinformation》2014,10(8):507-511
Endo- β-N-acetylgucosaminidases (ENGases) are the enzymes that catalyze both hydrolysis and
transglycosylation reactions. It is of interest to study ENGases because of their ability to synthesize glycopeptides.
Homology models of Human, Arabidopsis thaliana and Sorghum ENGases were developed and their active sites
marked based on information available from Arthrobacter protophormiae (PDB ID: 3FHQ) ENGase. Further, these
models were docked with the natural substrate GlcNAc-Asn and the inhibitor Man3GlcNAc-thiazoline. The catalytic
triad of Asn, Glu and Tyr (N171, E173 and Y205 of bacteria) were found to be conserved across the phyla. The crucial
Y299F mutation showing 3 times higher transglycosylation activity than in wild type Endo-A is known. The hydrolytic
activity remained unchanged in bacteria, while the transglycosylation activity increased. This Y to F change is found
to be naturally evolved and should be attributing higher transglycosylation rates in human and Arabidopsis thaliana
ENGases. Ligand interactions Ligplots revealed the interaction of amino acids with hydrophobic side chains and polar
uncharged side chain amino acids. Thus, structure based molecular model-ligand interactions provide insights into
the catalytic mechanism of ENGases and assist in the rational engineering of ENGases. 相似文献
173.
174.
The structure of a small collagenous fragment isolated from chicken hyaline cartilage 总被引:3,自引:0,他引:3
In previous experiments, two collagenous fragments were isolated from pepsin digests of chicken hyaline cartilage and called the high molecular weight, (HMW) and low molecular weight (LMW) fractions [3]. In the present experiments, the chains of LMW were isolated after denaturation and subsequent reduction and alkylation of interchain disulfide bridges and were further fractionated by carboxymethyl-cellulose chromatography. Four peaks were resolved during chromatography and were designated LMW 1, 2A, 2B, and 3. Amino acid analyses and peptide mapping after cleavage with trypsin, V8 protease, and cyanogen bromide showed that three genetically distinct chains must be present in LMW. Fractions 2A and 2B were very similar, but not identical, in structure. LMW 1, 2A plus 2B, and 3 were consistently isolated in approximately equal proportions, suggesting that the probable chain organization of LMW is [1][2A + 2B][3]. This suggestion was supported further by experiments that attempted to fractionate LMW by carboxymethyl-cellulose chromatography after denaturation but without reduction and alkylation of interchain disulfide bridges. No fractionation of LMW was achieved, the single peak subsequently being shown to contain LMW 1, 2A plus 2B, and 3. 相似文献
175.
The bacteriochlorophyll (P-800 and P-870) of the carotenoidless photoreaction center isolated from Rhodospirillum rubrum (strain G9) is bleached irreversibly when the preparations are exposed to intense near infrared light in the presence of oxygen. This effect is much smaller in preparations, extracted from the wild type, which contain, as shown earlier, 1 mol of spirilloxanthin per mol of P-870. This photodynamic effec is shown to be due to singlet O2. The oxidation of adrenaline in the presence of superoxide dismutase and the oxidation of 1,3-diphenylisobenzofuran are used as reporter reactions. Singlet oxygen is presumably generated by the triplet-triplet energy transfer 3bacteriochlorophyll → O2 (3Σ).Four purified bacterial carotenoids, spirilloxanthin, sphaeroidene, sphaeroidenone and chloroxanthin were attached onto the carotenoidless photoreaction center from strain G9 in nearly 1 : 1 mol ratios with respect to P-870. Once fixed, these carotenoids confer protection against the photodynamic bleaching of bacteriochlorophyll. The relative photoprotection efficiency was 1.0 for spirilloxanthin and sphaeroidene, 0.4 for chloroxanthin and 0.2 for sphaeroidenone. The fixed carotenoids display optical activity and their molar ellipticity appears to be correlated with their relative photoprotection efficiency. The efficiency of energy transfer to P-870 is 0.90 for sphaeroidene, 0.35 for sphaeroidenone, 0.30 for chloroxanthin and 0.20 for spirilloxanthin. The energy transfer efficiency from the carotenoids to bacteriochlorophyll is suggested to be governed by the rate of the internal conversion processes of the excited singlet state of the carotenoids.A study of the difference absorption and CD spectra of the reconstituted minus carotenoidless preparations leads to the interpretation that the fixed carotenoids are in a central monocis conformation. 相似文献
176.
Radiolabelling of DNA/polypeptide complexes in isolated bulk DNA and in residual nuclear matrix DNA by nick-translation 总被引:2,自引:0,他引:2
Conditions are described that allow 32P-radiolabelling and detection of tight complexes between DNA and polypeptides by nick-translation. Prolonged nick-translation of purified bulk DNA results in radiolabelled complexes migrating on SDS-polyacrylamide gels with apparent molecular weights of 68 kd and 54 kd respectively. Residual nuclear matrix DNA which is not accessible to DNase I on the nuclear level becomes accessible to radiolabelling by nick-translation on the nuclear matrix level. In this case the in situ radiolabelled complexes migrate on SDS-polyacrylamide gels with apparent molecular weights of 68 kd and 100 kd. The DNA/polypeptide complexes are stable during treatments with SDS, beta-mercapto ethanol and alkali which points to covalent bonds between the polypeptides and DNA strands. 相似文献