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David K. Grandy Richard Leduc Haripriya Makam Thomas Flanagan Emanuel J. Diliberto Jr. Olivier Civelli O. Humberto Viveros 《Cellular and molecular neurobiology》1992,12(2):185-192
1. A novel 1745-dalton pyroglutamyl peptide (BAM-1745)6 was recently isolated and characterized from bovine adrenal medulla chromaffin granules. Its amino acid sequence was found to be 93% identical to residues 580-593 of human chromogranin B (secretogranin I). 2. Based on this sequence a degenerate oligonucleotide probe was synthesized and used to identify a 2.4-kb bovine adrenal medulla chromogranin B cDNA. 3. The deduced polypeptide is 647 amino acids long and begins with a putative signal sequence of 20 residues as in the human, rat, and mouse proteins. Also conserved in the bovine protein is a tyrosine residue which may be sulfated, two N-terminal cysteines, and many paired basic amino acids which may serve as sites of posttranslational processing. The peptide BAM-1745 is flanked by paired basic amino acids and therefore is most likely a product of posttranslational processing. Bovine chromogranin B is 67, 58, and 58% identical to the human, rat, and mouse chromogranin B proteins, respectively. 4. The carboxyl terminus of bovine chromogranin B, including BAM-1745, was found to be the most conserved region of the polypeptide and may identify it as an important functional domain. 相似文献
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Visualization of poly(ADP-ribose) synthetase associated with polynucleosomes by immunoelectron microscopy 总被引:1,自引:0,他引:1
Antibodies showing a high specificity for poly(ADP ribose) synthetase have been purified. A fraction binding nonspecifically to histones present in antiserum and non-immune serum has been demonstrated by immunoblotting and separated by histone-Sepharose chromatography. The antibody without the nonspecific binding fraction was analyzed by Western blot with calf thymus protein extract and was found to react only with a band at 116 kDa. There was no reaction with purified topoisomerase I, this weak activity was copurified with poly(ADP-ribose) synthetase preparation. The specific IgG fraction has been used for the visualization of the interaction of poly(ADP-ribose) synthetase with chromatin by indirect gold-labelling. This immunomicroscopic study suggests that the synthetase is located in the inner part of polynucleosomes and would be associated preferentially with the core nucleosome. 相似文献
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An Electron Microscope Study of Intranuclear Inclusions in Mouse Liver and Hepatoma 总被引:8,自引:0,他引:8
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An electron microscope study of intranuclear inclusions which occur in giant cells in a transplantable mouse hepatoma and in enlarged liver cells in mice fed a diet containing bentonite demonstrates that these inclusions are formed by invaginations of the nuclear envelope, and corroborates a previous histochemical study which revealed that the contents of the inclusions are of cytoplasmic origin. In the hepatoma cells the intranuclear inclusions are abundant, small, and situated close to the border of the nucleus, and there are wide openings from the cytoplasm into the invaginations whose contents include lipid droplets, ergastoplasm, and structurally normal mitochondria. In the enlarged liver cells the inclusions are fewer in number, generally much larger than those in the hepatoma, hence they extend deeper into the nucleus, and the interior is continuous with the cytoplasm through only a small opening. Some normal ergastoplasm is present within the inclusions but all other constituents are abnormal. Both normal and degenerating mitochondria occur in the cytoplasm but only degenerating ones are found within the inclusions. Both types of inclusions arise in greatly enlarged cells in which an attempt is made to maintain the normal nuclear surface/nuclear volume ratio by the development of the invaginations of the nuclear envelope. 相似文献
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Fragmentation of bovine chromogranin A by plasma kallikrein 总被引:1,自引:0,他引:1
Chromogranin A has been reported to be processed in vivo by an as yet undefined proteinase(s) suggesting that it is a precursor of biologically active peptides such as pancreastatin. In this study, plasma kallikrein was used as a model proteinase to identify the cleavage sites exposed in bovine parathyroid chromogranin A. Purified bovine parathyroid chromogranin A was digested with human plasma kallikrein. The proteolytic fragments produced were isolated by HPLC and chemically characterized by amino acid composition and sequence analysis. The combined results indicate that the enzyme has preference for specific single Arg residues, cutting C-terminal to this amino acid, although certain pairs of basic sites were also cleaved. The characterized fragments were released in a selective manner from the whole molecule with rapid production of the fragments covering positions 1-247 and 352-358. 相似文献
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The fusarium mycotoxin zearalenone was transformed in cell suspension cultures of Zea mays giving α- and β-zearalenol and the β-D-glu cos ides of zearalenone and α- and β-zearalenol. The structure of zearalenone-4-β-D-glucopyranoside was determined by liquid — chromatography-mass spectrometry and specific hydrolysis with β-glucosidase. α- and β-zearalenol and their glucosides were identified by co chromatography using tic and HPLC and glucosidase — treatment Up to 50% of the mycotoxin added was bound to a non extractable or “bound” residue fraction. After treating this residue by a sequential cell wall fractionation procedure, zearalenone was found to be bound mainly to starch, hemicellulose, and lignin fractions. 相似文献
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Gene transfer is a major factor in bacterial evolution 总被引:17,自引:3,他引:14
Lateral gene transfer in four strains of Salmonella enterica has been
assessed using genomic subtraction. Strain LT2 (subspecies I serovar
Typhimurium) chromosomal DNA was used as target and subtracted by three
subspecies I strains of serovars Typhimurium (S21), Muenchen (S71), Typhi
(M229), and a subspecies V strain (M321). Data from probing random cosmids
of LT2 DNA with preparations of the residual LT2 DNA after subtraction were
used to estimate the amounts of LT2 DNA not able to hybridize to strains
S21, S71, M229, and M321 to be in the range of 84-106, 191-355, 305-629,
and 778-1,286 kb, respectively. Several lines of evidence indicate that
most of this DNA is from genes not present in strain M321 and not from
genes that have diverged in sequence. The amounts correlate with the
divergence of the four strains as revealed by multilocus enzyme
electrophoresis and sequence variation of housekeeping genes. Sequence of
39 of the fragments from the M321 subtracted residual LT2 DNA revealed only
six inserts of known gene function with evidence of both gain and loss of
genes during the development of S. enterica clones. Sixteen of the 39
segments have 45% or lower G+C content, below the species average, but over
half are within the normal range for the species. We conclude that even
within a species, clones may differ by up to 20% of chromosomal DNA,
indicating a major role for lateral transfer, and that on the basis of G+C
content, a significant proportion of the DNA is from distantly related
species.
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