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41.
42.
Modified hepatitis B virus surface antigen M protein particles (HBsAg M-P31c) produced in yeast is mainly composed of two differently glycosylated proteins, GP37 and GP34. GP37 has an N-linked sugar chain and O-linked sugar chains; and GP34 has an N-linked sugar chain bound to the peptide backbone P31. Although M-P31c vaccine elicits both anti-S and anti-pre-S2 antibodies, whether there are any differences between GP37 and GP34 in the ability to elicit these antibodies is still unknown. To clarify this issue, we prepared particles which were composed solely of GP37 or GP34 by affinity chromatography, using polymerized human serum albumin as a ligand and digestion with alpha-mannosidase. We also prepared particles composed solely of P31 by successive digestion with alpha-manosidase and endo-beta-N-acetylglycosaminidase H. The vaccines derived from these three kinds of particles elicited both anti-S and anti-pre-S2 antibodies in mice to the same extent as the original M-P31c vaccine. These results suggest that the N- and O-linked sugar chains of M-P31c component proteins produced in the host yeast cells have no effect on the ability to elicit anti-S and anti-pre-S2 antibodies and that there are no differences with respect to antibody response in mice between the two major components of M-P31c, GP37 and GP34. 相似文献
43.
The application of enzymatic transformations is attracting increasing attention. Until recently, such an approach has generally been confined to producing fine chemicals difficult to obtain through conventional chemical methods. Microbial nitrile hydratase (NHase) has now been applied to the industrial, kiloton-scale production of the important chemical commodity acrylamide. Recent progress in understanding microbial nitrile metabolism at both the gene and protein levels is permitting improvement of the acrylamide production process. 相似文献
44.
K Fujimoto I Kubota Y Yasuda-Kamatani H Minakata K Nomoto M Yoshida A Harada Y Muneoka M Kobayashi 《Biochemical and biophysical research communications》1991,177(2):847-853
Achatin-I previously purified from the ganglia of the African giant snail Achatina fulica was isolated from the atria of this snail. Achatin-I appeared to enhance the cardiac activity in two ways; centrally this peptide increased impulse frequency and produced spike broadening of the identified heart excitatory neuron, PON, and peripherally it enhanced amplitude and frequency of the heart beat. Achatin-I showed excitatory actions not only on the heart but on several other muscles. 相似文献
45.
Amyloplasts were isolated from a heterotrophic culture cell line of a woody plant, sycamore (Acer pseudoplatanus), and their DNA was purified. Conventional procedures for making a physical map were not easily applicable to the amyloplast DNA, since the yield of DNA was too low and the presence of repeated sequences interfered with the analysis. Therefore, the pieces of amyloplast DNA starting with a few micrograms of DNA were cloned in the lambda Fix vector, which is a derivative of lambda EMBL vectors improved for efficient cloning and gene walking. Cloned DNA fragments were randomly picked, mapped for restriction endonuclease sites by a refined procedure, and combined by overlapping their physical maps. The DNA library was also subjected to screening by gene walking using promoters recognized by T3 and T7 RNA polymerases in the vector to fill the gaps between sequences determined by overlapping the physical maps. In this way, we constructed the entire DNA library and the complete physical map of the amyloplast DNA. The sycamore amyloplast genome was composed of 141.7-kbp nucleotides with the same gene arrangement as that of tobacco chloroplasts. 相似文献
46.
The reactions of the NAD radical (NAD.) with ferric horseradish peroxidase and with compounds I and II were investigated by pulse radiolysis. NAD. reacted with the ferric enzyme and with compound I to form the ferrous enzyme and compound II with second-order rate constants of 8 X 10(8) and 1.5 X 10(8) M-1 s-1, respectively, at pH 7.0. In contrast, no reaction of NAD. with native compound II at pH 10.0 nor with diacetyldeutero-compound II at pH 5.0-8.0 could be detected. Other reducing species generated by pulse radiolysis, such as hydrated electron (eaq-), superoxide anion (O2-), and benzoate anion radical, could not reduce compound II of the enzyme to the ferric state, although the methylviologen radical reduced it. The results are discussed in relation to the mechanism of catalysis of the one-electron oxidation of substrates by peroxidase. 相似文献
47.
Structure of genomic DNA for rat platelet phospholipase A2 总被引:2,自引:0,他引:2
A genomic DNA for rat platelet phospholipase A2 was isolated by screening a rat genomic library with oligonucleotide probes based on its published amino acid sequence. The rat platelet phospholipase A2 gene had a total length of about 2.5 kb and contained five exons and four introns. The intron-exon structure of the rate gene was similar to that of human non-pancreatic phospholipase A2. 相似文献
48.
The reduction of oxy forms in cobalt-substituted hemoproteins by the hydrated electron (e(aq)-) was investigated by pulse radiolysis. The hydrated electron (e(aq)-) reacted with the oxy form of cobalt horseradish peroxidase (CoHRP) to form CoHRP. On the other hand, the initial product observed in the reaction of the oxy form of cobalt myoglobin (CoMb) with e(aq)- is neither CoMb nor Co3+ Mb. Subsequently, the product was found to convert to another form, the irreversible change in the porphyrin. In contrast to e(aq)-, both oxy forms of CoMb and CoHRP were reduced by various electron donors to form the cobaltic forms. 相似文献
49.
Native oxymyoglobin (MbO2) was isolated directly from the skeletal muscle of bigeye tuna (Thunnus obesus) with complete separation from metmyoglobin (metMb) on a CM-cellulose column. It was examined for its stability properties over a wide range of pH values (pH 5-12) in 0.1 M buffer at 25 degrees C. When compared with sperm whale MbO2 as a reference, the tuna MbO2 was found to be much more susceptible to autoxidation. Kinetic analysis has revealed that the rate constant for a nucleophilic displacement of O2- from MbO2 by an entering water molecule is 10-times higher than the corresponding value for sperm whale MbO2. The magnitude of the circular dichroism of bigeye tuna myoglobin at 222 nm was comparable to that of sperm whale myoglobin, but its hydropathy profile revealed the region corresponding to the distal side of the heme iron to be apparently less hydrophobic. The kinetic simulation also demonstrated that accessibility of the solvent water molecule to the heme pocket is clearly a key factor in the stability properties of the bound dioxygen. 相似文献
50.
T Ohtomo T Kobayashi Y Ohshima Y Usui M Suganuma K Yoshida 《Canadian journal of microbiology》1990,36(3):206-210
The interaction between the binding site of a polysaccharide (called compact colony forming active substance (CCFAS)), obtained from the cell surface of a strain of Staphylococcus, and human fibrinogen (HF) was investigated. The CCFAS was found to bind specifically to both the B beta and gamma chains of HF at pH 7.0 and 8.0, and the A alpha chain at pH 5.0. The binding of CCFAS with fibrinogen fragments obtained by digestion with plasmin were also investigated. Fragments with Mr of 55,000, 24,000, and 19,000 were the major bands precipitated by CCFAS at pH 7.0 and 8.0. Fragments with Mr of 85,000 and 75,000 bound to CCFAS at pH 5.0. Binding of CCFAS (7 micrograms) with fibrinogen could be inhibited by 1.2 micrograms of B beta chain and 1.5 micrograms gamma chain at alkaline pH or 6.2 micrograms of the A alpha chain at pH 5.0. CCFAS was, therefore, assumed to be specifically bonded with HF molecules, in the alkaline range at least, resulting in compact colony forming activity in serum soft agar and paracoagulation. 相似文献