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41.
R M Kamp Z J Yao B Wittmann-Liebold 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1983,364(2):141-155
Tryptic peptides of the ribosomal proteins S11, L9 and L29 were separated by reversed phase chromatography under conditions which enabled direct micro-sequencing with the 4-(dimethylamino)azobenzene-4'-isothiocyanate/phenylisothiocyanate double coupling method [Chang, Brauer, Wittmann-Liebold (1978) FEBS Lett. 93, 205-214]. The peptides were separated on a RP-18 column employing volatile buffers at pH 2.0, 4.1 and 7.8. Depending on the different chromatographic behaviour of the peptide mixture, the elution gradient was optimised for each hydrolysate using 20 micrograms of the hydrolysed protein. Preparative separations were made with 150-250 micrograms. At least 80% of the peptides could be isolated by these techniques and used for direct micro-sequencing without further purification or desalting. The results show that the high-performance liquid chromatographic method employed allows easy isolation and sequencing with minute amounts of peptides. 相似文献
42.
Eleonora Condrea Bruce E. Rapuano Karen R. Soons Philip Rosenberg 《Life sciences》1983,32(13):1455-1461
The effects on some pharmacological and enzymatic properties were determined following methylation of histidine at the enzymatic active site of the basic relatively toxic and the acidic relatively non-toxic phospholipases A2. Following methylation a very low residual enzymatic activity (0.4 -- 1% of control) was accompanied by a parallel loss in intraventricular lethality, anticoagulant potency, direct hemolytic action and ability to block directly and indirectly evoked contractions of the mouse phrenic nerve-diaphragm preparation. Since methylation does not impair the enzyme's ability to bind monomeric of micellar substrates or Ca2+, the results suggest that the pharmacologicallly active region of the molecule is different from the micellular substrate binding site but strongly influenced by the invariant histidine-48 located at the enzymatic active site. 相似文献
43.
Recessive lethal mutations in the T/t-complex of the mouse characteristically show defective genetic complementation, even when they affect very different stages of embryogenesis and are known to be nonallelic. To address the question of their genetic or functional relationship, we have applied the cis-trans test, using several well defined recombinant t-chromosomes that carry two or more lethal mutations, and others that are devoid of specific lethals. We show here that the defective complementation that occurs between different t-lethals is a specific result of the trans configuration; thus these genes, which may map as much as 15 cM apart, constitute a functional unit. Some speculations are presented to interpret this enigma in terms of DNA plasticity. 相似文献
44.
Olga O. Blumenfeld Anthony M. Adamany Karen V. Puglia Wladyslaw W. Socha 《Biochemical genetics》1983,21(3-4):333-348
Chimpanzee erythrocytes express strong M but weak, occasional N blood-group activity, as detected by anti-M and anti-N reagents. We have found that the M activity is carried by a major membrane glycoprotein that is similar but not identical to the human MM glycoprotein (glycophorin A). We have isolated and characterized this glycoprotein from erythrocyte membranes of four individual chimpanzees. The purified glycoproteins strongly inhibited agglutination of M cells by rabbit anti-human M sera and only weakly inhibited the agglutination of N cells by rabbit anti-human N sera. They also displayed medium-to-strong inhibitory activity against chimpanzee iso- and crossimmune antisera tested with chimpanzee erythrocytes of various V-A-B-D and Wc specificities, which are known as chimpanzee extensions of the human type M-N system and the Miltenberger counterpart, respectively. Each glycoprotein was cleaved with CNBr into three fragments, whose size, solubility, and composition were analogous to those obtained by similar treatment of the human M-N antigens. The amino-terminal fragment was found to be a glycooctapeptide whose amino acid composition and partial sequence indicated that it is an intermediate form of the human M and N glycooctapeptides. Its carbohydrate content comprised two threonine-linked saccharide units that, although similar in composition to the human threonine-linked units, were fewer in number than the three units found in the corresponding human glycooctapeptides. Structural similarities to the human antigens strongly suggest that the amino terminus bears the major antigenic determinants of the molecule, and the occurrence in this region of numerous, albeit rare, variants among humans and in chimpanzees indicates that the corresponding coding sequence of the structural gene is particularly susceptible to mutational events. We conclude that the chimpanzee M gene product is a variant of the human type and that the chimpanzee gene is an allele of the human polymorphic M-N locus.This research was supported by National Institutes of Health Grants GM 16389 and HL 19011 and March of Dimes Grant 1-661. 相似文献
45.
Howard R. Morris Karen E. Batley Nigel G. L. Harding Richard A. Bjur John G. Dann Rodney W. King 《The Biochemical journal》1974,137(2):409-411
An elastase digest of a protein of unknown structure, dihydrofolate reductase, was studied by mass spectrometry. This soluble digest contained a large number of small peptides in different yields, within the ideal molecular-weight range (200-1200) for mixture-analysis mass spectrometry. Sequences of the major component peptides in the digest are reported. 相似文献
46.
14C-Sterigmatocystin isolated from cultures of supplemented with (1-14C)acetate was shown to be efficiently converted to aflatoxin B1 by the resting mycelium of . The experimental results may indicate a biosynthetic pathway leading from 5-hydroxysterigmatocystin to sterigmatocystin and then to aflatoxin B1. 相似文献
47.
Comparison of α1 -Adrenergic Receptor-Stimulated Inositol Phosphate Formation in Primary Neuronal and Glial Cultures 总被引:3,自引:3,他引:0
alpha 1-Adrenergic receptor binding sites and norepinephrine-stimulated 3H-inositol phosphate (3H-InsP) accumulation were measured in primary cultures of neurons and glia from 1-day-old rat brains. The density of alpha 1-adrenergic receptor binding sites was approximately three times higher in membranes from neurons compared to glia. Although norepinephrine was slightly more potent in stimulating 3H-InsP formation in neurons than in glia, the maximal response was greater in glial cells. Norepinephrine-stimulated 3H-InsP formation remained constant for [3H]inositol prelabelling periods of 1-14 days in neurons, whereas the response increased with time in glia and was maximal after 7-10 days of prelabelling. Both the incorporation of [3H]inositol into lipid and basal levels of 3H-InsPs were lower in glial cells than in neurons, which accounted for the greater percent stimulation in glia. Pretreatment with phenoxybenzamine decreased norepinephrine-stimulated 3H-InsP formation in a dose-dependent manner in both neurons and glia by decreasing the maximal response without altering potency. HPLC separation showed that similar types of 3H-InsPs were accumulated in neurons and glial cells. These results demonstrate that alpha 1-adrenergic receptors exist on both neurons and glial cells and activate 3H-InsP accumulation in both cell types. Although receptor density is higher in neurons than in glia, the 3H-InsP response is higher in glia. This difference does not appear to be due to different receptor reserves, but may be due to differential coupling mechanisms in the two cell types. 相似文献
48.
Clare M. Baecher Karen S. Dorfman Marie-Geneviève Mattei John G. Frelinger 《Immunogenetics》1990,31(5-6):307-314
Mouse leukosialin, previously known as the 3E8 antigen, is expressed primarily on cells of the hematopoietic and lymphoid lineages and is shown to be the mouse homologue to the human leukosialin/sialophorin and rat W3/13 molecules. A partial leukosialin cDNA clone was isolated via cross-species hybridization with a portion of a human leukosialin cDNA. This mouse cDNA clone was used to demonstrate that the leukosialin isoforms are encoded by a single mRNA species of approximately 4.2 kilobases (kb) and that the leukosialin gene is located on chromosome 7. Based on these results, mouse leukosialin is given the designation Ly48.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number M30693. 相似文献
49.
50.
Fetal cells in maternal blood: recovery by charge flow separation 总被引:11,自引:0,他引:11
S. S. Wachtel David Sammons Michael Manley Gwendolyn Wachtel Garland Twitty Joseph Utermohlen Owen P. Phillips Lee P. Shulman Douglas J. Taron U. R. Müller Peter Koeppen Teresa M. Ruffalo Karen Addis Richard Porreco Joyce Murata-Collins Natalie B. Parker Loris McGavran 《Human genetics》1996,98(2):162-166
Fetal blood cells can be recovered from the maternal circulation by charge flow separation (CFS), a method that obviates the
risks associated with amniocentesis and chorionic villus sampling. By CFS, we processed blood samples from 13 women carrying
male fetuses, 2 carrying fetuses with trisomy 21, and 1 who had delivered a stillborn infant with trisomy 18. On average more
than 2000 fetal nucleated red blood cells were recovered per 20-ml sample of maternal blood. Recovery of fetal cells was confirmed
by fluorescence in situ hybridization with probes for chromosomes Y, 18 and 21. After culturing of CFS-processed cells, amplification
by the polymerase chain reaction revealed Y-chromosomal DNA in clones from four of six women bearing male fetuses, but not
in clones from three women bearing female fetuses.
Received: 8 January 1996 / Revised: 22 March 1996 相似文献