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91.
Hostinová E Solovicová A Dvorský R Gasperík J 《Archives of biochemistry and biophysics》2003,411(2):189-195
Raw-starch-degrading glucoamylases have been known as multidomain enzymes consisting of a catalytic domain connected to a starch-binding domain (SBD) by an O-glycosylated linker region. A molecular genetics approach has been chosen to find structural differences between two related glucoamylases, raw-starch-degrading Glm and nondegrading Glu, from the yeasts Saccharomycopsis fibuligera IFO 0111 and HUT 7212, respectively. We have found that Glm and Glu show a high primary (77%) and tertiary structure similarity. Glm, although possessing a good ability for raw starch degradation, did not show consensus amino acid residues to any SBD found in glucoamylases or other amylolytic enzymes. Raw starch binding and digestion by Glm must thus depend on the existence of a site(s) lying within the intact protein which lacks a separate SBD. The enzyme represents a structurally new type of raw-starch-degrading glucoamylase. 相似文献
92.
A critical event in eukaryotic DNA replication involves association of minichromosome maintenance (MCM2-7) proteins with origins, to form prereplicative complexes (pre-RCs) that are competent for initiation. The ability of mutants defective in MCM2-7 function to complete meiosis had suggested that pre-RC components could be irrelevant to premeiotic S phase. We show here that MCM2-7 proteins bind to chromatin in fission yeast cells preparing for meiosis and during premeiotic S phase in a manner suggesting they in fact are required for DNA replication in the meiotic cycle. This is confirmed by analysis of a degron mcm4 mutant, which cannot carry out premeiotic DNA replication. Later in meiosis, Mcm4 chromatin association is blocked between meiotic nuclear divisions, presumably accounting for the absence of a second round of DNA replication. Together, these results emphasize similarity between replication mechanisms in mitotic and meiotic cell cycles. 相似文献
93.
High-performance liquid chromatographic analysis and separation of N-feruloylserotonin isomers 总被引:2,自引:0,他引:2
Pavlík M Laudová V Grüner K Vokác K Harmatha J 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2002,770(1-2):291-295
The N-feruloylserotonin containing fraction was isolated from seeds of Leuzea carthamoides (Willd.) DC by solvent extraction followed by column chromatography on silica gel or on Sephadex LH-20. Nuclear magnetic resonance spectroscopic analysis of the isolated fraction showed the presence of four structurally related compounds. These compounds were identified as four isomers of N-feruloylserotonin: N-(Z)-feruloylserotonin, N-(Z)-isoferuloylserotonin, N-(E)-feruloylserotonin and N-(E)-isoferuloylserotonin. They were analyzed by HPLC on Separon SGX C18, Separon SGX and Separon SGX phenyl, using various mobile phases. Separon SGX phenyl phase was found the most efficient for a rapid analysis and for the final separation of the N-feruloylserotonin isomers. 相似文献
94.
95.
James M. Foppoli Lean Kuan Ch'ng Albert A. Benedict Juraj Ivanyi Josef Derka Edward K. Wakeland 《Immunogenetics》1979,8(1):385-404
Based on a survey of 36 inbred and 8 partially inbred chicken lines and outbred jungle fowl, and with 29 alloantisera generated in different laboratories, 13 7S Ig and 5 IgM allotypes were designated and a new system of nomenclature for chicken Ig polymorphisms was developed. The survey also revealed considerable genetic polymorphism in the structural gene(s) (G-1) responsible for the production of 7S Ig H chains. IgM H chains, encoded by theM-1 locus were less polymorphic. NineG-1 and fourM-1 gene alleles were delineated in highly inbred lines by the formation of unique combinations ofG-1 orM-1 specificities. Five additionalG-1 alleles were found in chicken lines and jungle fowl segregating for allotypes. Thirty-three percent of theG-1M-1 haplotypes theoretically expected, were detected in inbred lines. 相似文献
96.
High-level expression and purification of a recombinant hBD-1 fused to LMM protein in Escherichia coli 总被引:9,自引:0,他引:9
In this work, we present the production of an active 43 aa recombinant human beta-defensin-1 (rhBD-1(43)) in Escherichia coli AD202 cells using specific pLMM1-rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with isolation procedures and toxicity caused by rhBD-1 to the host organism. As a result, the majority of fusion protein (LMM-rhBD-1(43)) was obtained in the soluble state, isolated by a low salt-high salt treatment of total cell protein. The rhBD-1(43) was cleaved from the fusion with Protease 4 and purified on CM Sepharose Fast Flow column with the yield of approximately 1 mg rhBD-1(43) from 6 g of wet weight cells. Purified rhBD-1(43) showed antimicrobial activity against E. coli ML-35p at a concentration of 129 microM. The procedure of rhBD-1 expression and purification we present can provide a reliable and simple method for production of different cationic peptides for biological studies. 相似文献
97.
Fuyi Wang Juraj Bella John A. Parkinson Peter J. Sadler 《Journal of biological inorganic chemistry》2005,10(2):147-155
The ruthenium arene anticancer complex [(6-bip)Ru(en)Cl][PF6] (1) (bip is biphenyl, en is ethylenediamine) reacted slowly with the amino acid L-histidine (L-His) in aqueous solution at 310 K. Two L-His adducts of 1 were separated by high-performance liquid chromatography and identified by electrospray ionisation mass spectrometry and NMR: an imidazole N-bound complex [(6-bip)Ru(en)(N–L-His)]2+, and an N-bound complex [(6-bip)Ru(en)(N–L-His)]2+. At 310 K, after 24 h only about 22% of complex 1 (2 mM) reacted with L-His, and of the unreacted 1, 59% had hydrolysed. In the presence of 100 mM NaCl, approximately 90% of 1 remained unreacted. In aqueous solution or triethylammonium acetate (TEAA) buffer (pH 7.6), 15N-labelled 1 reacted with cytochrome c to give two monoruthenated protein adducts. The reaction reached equilibrium within 2 h by which time approximately 50% of cytochrome c was ruthenated. On the basis of [1H, 15N] NMR data, one adduct may have Ru bound to the N-terminus, and the other to a carboxylate group on the protein. In TEAA buffer and at 310 K, more than 90% of the 14-mer oligonucleotide d(TATGTACCATGTAT) reacted with 2 mol Eq of 1 to give rise to monoruthenated and diruthenated oligonucleotide adducts. The presence of cytochrome c (1 mol Eq) or L-His (4 mol Eq) had little effect on the course of the reaction with the oligonucleotide. In cells, DNA (or RNA) may be a favoured reaction site for this Ru anticancer complex.Electronic supplementary material is available for this article at .
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98.
Xiaowen?Yang Juraj?Gregan Karola?Lindner Hedi?Young Stephen?E?KearseyEmail author 《BMC molecular biology》2005,6(1):13
Background
Cdc23/Mcm10 is required for the initiation and elongation steps of DNA replication but its biochemical function is unclear. Here, we probe its function using a novel approach in fission yeast, involving Cdc23 cleavage by the TEV protease. 相似文献99.
Gordon A. Morris Zdenka Hromdkov Anna Ebringerov Anna Malovíkov Juraj Alfldi Stephen E. Harding 《Carbohydrate polymers》2002,48(4):351-359
Citrus pectin with a low degree of methyl esterification (LMP) and its deesterified form, potassium pectate (KP), were modified with a low amount of UV-absorbing substituents. For this purpose, two different substitution reactions were used (a) alkylation of hydroxyl groups with p-carboxybenzyl bromide in aqueous alkali and (b) alkylation of the carboxylate group with benzyl bromide in the DMSO/TBAI/catalyst system. Chemical and spectroscopic methods reveal a low degree of substitution (DS<0.1) for the derivatives. The hydrodynamic properties were assessed by analytical ultracentrifugion, viscometry, and HPGPC. The results indicate that the introduction of small amounts of p-carboxybenzyl ether groups practically had no effect on the hydrodynamic properties in the case of KP, whereas, it was accompanied with a decrease of the molecular mass for LMP. The degradation was more pronounced during the benzyl esterification of LMP. The results confirmed that LMP is susceptible to chain cleavage due to β-elimination during both modification reactions. However, KP seems to be more tolerant of the reaction conditions. 相似文献
100.
Insect feeding deterrent activity of bisabolangelone and of some sesquiterpenes of eremophilane type
The insect feeding deterrent activity of some sesquiterpenes of furanoeremophilane type and of the related eremophilanolide type were tested towards three selected storage pests: Sitophilus granarius, Tribolium confusum and Trogoderma granarium. The results were compared with the biogenetically related compound of bakkenolide type, known as a potent antifeedant. Bisabolangelone, tested on the same insects, exhibited the strongest activity, and may be included in the class of very good insect feeding deterrents. The antifeeding activity of bisabolangelone and bakkenolide A towards larvae of Leptinotarsa decemlineata was also observed. 相似文献