共查询到20条相似文献,搜索用时 15 毫秒
1.
G Alcaraz J Marti D Moinier M Fougereau 《Biochemical and biophysical research communications》1981,99(1):30-36
Calf fetuin, one of the 3 major known fetal proteins has been isolated by a two-step purification procedure and characterized by aminoacid composition. The purified glycoprotein, which consisted of a single chain, was submitted to 47 steps of automatic aminoacid sequencing, allowing to determine 44 positions. This section of the molecule was devoid of carbohydrates. Comparison of this sequence with a variety of detectable potentially related protein did not allow to point to any detectable homology. 相似文献
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S Shoji A Tashiro K Furuishi O Takenaka Y Kida S Horiuchi T Funakoshi Y Kubota 《Biochemical and biophysical research communications》1989,162(2):724-732
Novel antibodies were raised against a synthetic NH2-terminal myristoyl glycine moiety which is characteristic of N-myristoyl-proteins. Antisera raised against N-myristoyl-Gly-hemocyanin reacted with N-myristoyl-Gly-[125I]albumin. The immunoreaction was competed for by albumin conjugated with N-myristoyl-glycine, while underivatized albumin had no effect. Of the [3H]myristate-labeled proteins detected, pp60v-src, which is a transforming protein of Rous sarcoma virus, and p19gag and p17gag, which are core proteins in the human T-cell leukemia virus and the human immunodeficiency virus, were identified as N-myristoylated proteins by the radioimmunoprecipitation analyses with the antibody. 相似文献
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NH2-terminal processing of Bacillus subtilis alpha-amylase 总被引:4,自引:0,他引:4
Mature alpha-amylase of Bacillus subtilis is known to be formed from its precursor by removal of the NH2-terminal 41-amino acid sequence. To study the mechanism of this processing, the extracellular forms of alpha-amylase were analyzed for B. subtilis N7 alpha-amylase cloned and expressed in B. subtilis. The major form (form N34) isolated from log phase cultures in L-broth had an NH2 terminus corresponding to position 34 from the initiator Met but appeared to be microheterogeneous, as judged by native gel electrophoresis. The major forms from stationary phase cultures had NH2 termini at positions 40 (form N40) or 42 (form N42) and were homogeneous. The conversion of the larger to smaller forms could be achieved in culture supernatants or partially purified samples. The process N34----N40 was inhibited by EDTA; N40----N42 was facilitated by Ca2+. Phenylmethylsulfonyl fluoride inhibited the former but not the latter process. These results suggest that the signal peptidase cleavage site 30 decreases 35 is -Ala-Ala-Ala-Ser-Ala-Glu-Thr- (arrow or further upstream) and that proteolytic maturation occurs after secretion, which involves at least two different processing enzymes. 相似文献
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Ovalbumin utilizes an NH2-terminal signal sequence 总被引:7,自引:0,他引:7
In synchronized translation experiments in the wheat germ and reticulocyte lysate systems, ovalbumin (385 amino acids) was glycosylated by and segregated in dog pancreatic microsomes only if microsomes were added before the nascent ovalbumin polypeptide contained less than 150 amino acids. This would place the "signal" sequence of ovalbumin prior to residue 150, in contrast to a previous report. 相似文献
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R. Håkanson F. Sundler A. Nobin N. -O. Sjöberg L. Edvinsson L. -I. Larsson 《Cell and tissue research》1974,150(2):281-290
Summary In the mammalian pituitary formaldehyde-ozone treatment induces strong fluorescence in the cells of the pars intermedia and moderate to strong fluorescence in numerous cells of the pars distalis. Maximum excitation is at 370–375 nm and maximum emission at 495–505 nm. The properties of the cellular fluorescence are indistinguishable from those of tryptamine or peptides with NH2-terminal tryptophan. From chemical analysis such peptides seem to occur abundantly in the mammalian pituitary. The concentration of these peptides agrees very well with the number and fluorescence intensity of the cells in all species studied. Furthermore, the tryptophyl peptides in the various parts of the pig pituitary have a distribution quite parallel to that of the fluorescent cells. As we have failed to detect tryptamine in the pituitary, we conclude that the formaldehyde-ozone-induced fluorescence in the adenohypophysis reflects the presence of tryptophyl peptides.This study was supported by grants from the Swedish Medical Research Council (04X-1007; 04X-3764), the Ford Foundation, Harald and Greta Jeanssons stiftelse and Riksföreningen mot Cancer (660-K73-01X).For brevity occasionally referred to as tryptophyl peptides. 相似文献
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The NH2-terminal residues of Bacillus subtilis proteins 总被引:4,自引:0,他引:4
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The concentrations and molecular forms of urinary and plasma gastrin from normal subjects were studied by radioimmunoassays using two region-specific antisera. Urinary concentration of NH2-terminal big gastrin (G-34) immunoreactivity was several hundred times as great as that of COOH-terminal gastrin immunoreactivity. Fractionation of urine extract showed a broad giant peak of NH2-terminal G-34 immunoreactivity (gastrin fragments "U") eluting in a later position than G-34(1-17) by Sephadex G-50 column chromatography. HPLC revealed that urinary NH2-terminal G-34 immunoreactivity was composed of four fragments including G-34(1-8), G-34(1-9), and G-34(1-10). Sephadex G-50 column chromatography of plasma extract revealed two or three peaks of NH2-terminal G-34 immunoreactivity, and a major peak eluted in the same position as urinary gastrin fragments "U". These results and data on renal clearances suggest that most of all gastrin fragments "U" in plasma are excreted in urine without renal reabsorption, whereas almost all of plasma COOH-terminal gastrin peptides including G-34 and little gastrin (G-17) are removed and metabolized in the kidney. 相似文献
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Abstract— The NH2-terminal amino acids of Wolfgram and Folch-Lees proteolipids of bovine and human CNS myelin were determined using the cyanate method (Starke & Smyth , 1963) followed by direct amino acid analysis of the products. Glycine predominated in every case and was recovered in amounts similar to the results described by Whikehart & Lees (1973), who used a dansylation technique followed by thin layer chromatography of the DNS-amino acids. In the present study substantial amounts of glutamic acid, serine, alanine and aspartic acid were also recovered, plus traces of other amino acids. Few differences were observed between Wolfgram and Folch-Lees proteolipids. The end group products of purified W1 proteolipid of bovine Wolfgram fraction, of diazometholysed Folch-Lees proteolipid, and of a sample of phosphatidyl serine had essentially the same composition. The similarity of these results, especially for both fractionated and unfractionated Wolfgram proteolipid, may be evidence that the observed products are derived from phosphoglycerides present in proteolipid rather than from the actual NH2-terminals of the protein. 相似文献
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In many voltage-gated K(+) channels, N-type inactivation significantly accelerates the onset of C-type inactivation, but effects on recovery from inactivation are small or absent. We have exploited the Na(+) permeability of C-type-inactivated K(+) channels to characterize a strong interaction between the inactivation peptide of Kv1.4 and the C-type-inactivated state of Kv1.4 and Kv1.5. The presence of the Kv1.4 inactivation peptide results in a slower decay of the Na(+) tail currents normally observed through C-type-inactivated channels, an effective blockade of the peak Na(+) tail current, and also a delay of the peak tail current. These effects are mimicked by addition of quaternary ammonium ions to the pipette-filling solution. These observations support a common mechanism of action of the inactivation peptide and intracellular quaternary ammonium ions, and also demonstrate that the Kv channel inner vestibule is cytosolically exposed before and after the onset of C-type inactivation. We have also examined the process of N-type inactivation under conditions where C-type inactivation is removed, to compare the interaction of the inactivation peptide with open and C-type-inactivated channels. In C-type-deficient forms of Kv1.4 or Kv1.5 channels, the Kv1.4 inactivation ball behaves like an open channel blocker, and the resultant slowing of deactivation tail currents is considerably weaker than observed in C-type-inactivated channels. We present a kinetic model that duplicates the effects of the inactivation peptide on the slow Na(+) tail of C-type-inactivated channels. Stable binding between the inactivation peptide and the C-type-inactivated state results in slower current decay, and a reduction of the Na(+) tail current magnitude, due to slower transition of channels through the Na(+)-permeable states traversed during recovery from inactivation. 相似文献
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NH2-terminal processing of actin in mouse L-cells in vivo 总被引:1,自引:0,他引:1
When Dictyostellium discoideum actin is synthesized in vitro, it is made as a 43,000-dalton polypeptide with an NH2-terminal N-acetylmethionine. The acetylmethionine is then cleaved post-translationally, and the new NH2-terminal aspartic acid is acetylated to give the mature form of actin. Inhibition of methionine acetylation prevents methionine cleavage (Redman, K., and Rubenstein, P. (1981) J. Biol. Chem. 256, 13226-13229). In this paper, we describe the results of experiments designed to discover whether this novel actin processing pathway is peculiar to the rabbit reticulocyte lysate system or whether it is utilized by mammalian cells in vivo as well. We show that in mouse L-929 cells, actin is made as a 43,000-dalton protein with an NH2-terminal N-acylmethionine residue. Experiments using thin layer chromatography and digestion of the acylmethionine residue with hog kidney acylase I demonstrate that the acyl group is an acetyl residue. Pulse-chase experiments show that over the course of 1 h, this precursor is transformed first to an actin with a free NH2-terminal aspartic acid and is subsequently converted to mature L-cell actin with an acetylaspartic acid NH2 terminus. The half-life of the initial actin precursor in the cell appears to be approximately 12-15 min. These studies demonstrate the existence of this novel actin processing pathway in vivo and suggest that it is used for those actins where, in the gene, the initiator methionine codon directly precedes the codon for aspartic or glutamic acids, the residues normally found at the actin NH2 terminus. 相似文献
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F Gümü?el R H Cool A Weijland P H Anborgh A Parmeggiani 《Biochimica et biophysica acta》1990,1050(1-3):215-221
Mutagenesis was carried out in the N-terminal domain of elongation factor Tu (EF-Tu) to characterize the structure-function relationships of this model GTP binding protein with respect to stability, the interaction with GTP and GDP, and the catalytic activity. The substitutions were introduced in elements around the guanine nucleotide binding site or in the loops defining this site, in the intact molecule or in the isolated N-terminal domain (G domain). The double substitution Val88----Asp and Leu121----Lys, two residues situated on two vicinal alpha-helices, influences the basic activities of the truncated factor to a limited extent, probably via long-range interactions, and induces a destabilisation of the G domain structure. The functional alterations brought about by substitutions on the consensus sequences 18-24 and 80-83 highlight the importance of these residues for the interaction with GTP/GDP and the GTPase activity. Mutations concerning residues interacting with the guanine base lead to proteins in large part insoluble and inactive. In one case, the mutated protein (EF-TuAsn135----Asp) inhibited the growth of the host cell. This demonstrates the crucial role of the base specificity for the active conformation of EF-Tu. The obtained results are discussed in the light of the three-dimensional structure of EF-Tu. 相似文献
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H Takakura S Tsunasawa M Miyagi J R Warner 《The Journal of biological chemistry》1992,267(8):5442-5445
Using a mutant of Saccharomyces cerevisiae defective in the NAT1 gene, that encodes one of the NH2-terminal acetyltransferases, we have identified 14 ribosomal proteins whose electrophoretic mobility at pH 5.0 suggests they carry an additional charge, presumably due to the lack of NH2-terminal acetylation. At least 30 other ribosomal proteins from the mutant are electrophoretically normal. Attempted NH2-terminal analysis of most of the presumed acetylated proteins from wild type cells indicated that all were blocked. NH2-terminal analysis of the same proteins from the nat1 mutant strain yielded unique sequences. Each one carries an NH2-terminal serine. We conclude that these are normally acetylated due to the presence of the NAT1 gene product. It seems surprising that cells whose ribosomes have been altered to this degree grow rather well and synthesize the same spectrum of proteins as do wild type cells (Mullen, J. R., Kayne, P. S., Moerschell, R. P., Tsunasawa, S. Gribskov, M., Sherman, F., and Sternglanz, R. (1989) EMBO J. 8, 2067-2075). Finally, this analysis has provided the first sequence information available for several of the acetylated ribosomal proteins and for one non-acetylated ribosomal protein, which is clearly the product of the MFT1 gene (Garrett, J. M., Singh, K. K., Vonder Haar, R. A., and Emr. S. D. (1991) Mol. Gen. Gen. 225, 483-491). 相似文献
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T M Pozdniakova V N Rybalchuk A V Il'ina Iu A Davidovich S V Rogozhin 《Ukrainski? biokhimicheski? zhurnal》1986,58(2):10-15
Peptides Gly-Pro-Arg-Pro and Gly-His-Arg-Pro (fibrin alpha- and beta-chain NH2-terminal analogs, respectively) are studied for their effect on fibrinogen (F) and fibrin NH2-terminal disulphide knot (N-DSK) specific binding. Both peptides are found to inhibit the formation of soluble and insoluble F-N-DSK-complexes through inhibition of the interdomain D-E-binding. Gly-Pro-Arg-Pro is much more potent inhibitor than Gly-His-Arg-Pro. Lowering the insoluble F-N-DSK-copolymer quantity by concentration-dependent way these peptides do not change its composition described by the formula [F(N-DSK)2]n. Invariability of fibrinogen and N-DSK copolymer structure is asserted. In this structure neighbouring fibrinogen molecules are bound by two N-DSK molecules via the D1-E1 and D2-E2 binding sites. 相似文献
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V N?thig-Laslo 《Biochimica et biophysica acta》1986,874(3):312-317
In order to explore fully how ligand- and temperature-induced alterations in the spin states of heme iron are related to protein readjustments, the spin label 4-isothiocyanate (I) was covalently attached at beta-93 cysteines and at NH2-terminal valines of various heme-iron ligand forms of human hemoglobin. It was found that the mobility of NH2-terminally bound spin labels depends on the magnetic moment of the heme iron. There is a an approximately linear relationship between the magnetic moment of the heme iron and the mobility of NH2-terminally bound spin labels. In accordance with our previous results, the temperature dependence of ESR spectra of spin-labeled hemoglobin suggests the temperature-induced protein conformational change in those heme-iron ligand forms that are characterized by the equilibrium of the spin states of the heme iron. The conformational change was sensed at both spin-label-binding sites: at beta-93 cysteines and at NH2-terminal valines. 相似文献