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1.
Two crystal forms of lysozyme from the egg white of the embden goose (Anser anser) have been obtained, both of which are suitable for X-ray diffraction analysis. The monoclinic form has space group P21 with cell dimensions , β = 116 ° and the triclinic form (space group P1) has cell dimensions of and α = 98.8 °, β = 102.5 °, γ = 90.5 °. 相似文献
2.
Ilkka Kouvonen 《生物化学与生物物理学报:生物膜》1981,646(2):268-274
The pig intestinal intrinsic factor receptor has been isolated and dissociated into its α and β subunits. The β subunit was found to be more hydrophobic than the α subunit. In a detergent solution only the α subunit was accessible to digestion with papain. The whole isolated receptor was introduced into artificial single bilayer liposomes where is apparently was randomly oriented. Liposomes containing the receptor were digested with papain and the polypeptide segments that stayed in the lipid fraction were extracted and analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Four species were found with values of 23 000, 45 000, 70 000 and 86 000. 相似文献
3.
Raman K. Roy James D. Potter Satyapriya Sarkar 《Biochemical and biophysical research communications》1976,70(1):28-36
The Ca2+-regulatory tropomyosin-troponin complex was purified from chick embryonic muscles by a combination of DEAE-cellulose chromatography and (NH4)2SO4 fractionation. The embryonic complex was very similar to that obtained from adult chicken muscles with respect to stoichiometry of components and biological activity. Tropomyosin of embryonic skeletal muscles contains both α and β subunits, the β form being the major species. In the adult stage the β form is decreased with a concomitant increase in the α form. These results indicate that i) the Ca2+-regulatory proteins are not deficient in early embryonic muscles as previously thought (Hitchcock, S.E., Develop. Biol. , 399, 1970), and ii) different structural genes coding for tropomyosin subunits are expressed differentially in embryonic and adult muscle fibers. 相似文献
4.
Antonio Parets Soler Louise Reibel Georges Schapira 《Biochemical and biophysical research communications》1981,99(4):1427-1432
The translation of rabbit α globin mRNA in a Krebs II ascites cellfree system was more dependent upon the K+ concentration than rabbit β globin mRNA. The optimal KCl concentration was approximately 70 mM for the synthesis of the α chain and between 80 and 90 mM for that of the β chain. With CH3 CO2K the optimum concentration for α chain synthesis was also 70 mM but the optimum for the β chain synthesis was not sharp any more and ranged from 70 mM to over 110 mM. In the range of the optimal Mg2+ concentration for the α and β globin chain synthesis the ratio decreased when the Mg2+ concentration increased. In the presence of DTT and EDTA the optimal KCl concentration for both α and β globin chain synthesis decreased. 相似文献
5.
D2O-alanine exchange reactions catalyzed by alanine racemase and glutamic pyruvic transminase 总被引:1,自引:0,他引:1
NMR studies in D2O (>90%) reveal that Alanine Racemase (5.1.1.1.) from catalyzes the exchange of the α hydrogen of - and -alanine with D2O. Glutamic Pyruvic Transaminase (2.6.1.2.) and Glutamic Oxaloacetic Transaminase (2.6.1.1.) catalyze the exchange of α and β hydrogens of -alanine. The rates of exchange of α and β hydrogens appear to be of the same order of magnitude. The transaminase catalyzed exchange is enhanced by catalytic amounts of pyruvate. The side chain of -alanine is held more rigidly at the active site of transaminase so that the planar conjugated system can be extended to include the α and β carbons. A generalized mechanism is proposed for the action of pyridoxal phosphate dependent transaminases which extends Braunstein and Snell mechanism to include the structures which contribute to the labilization of β hydrogens of amino acids by the transaminases that have been studied. 相似文献
6.
The immunologic cross-reactivity of the α and α+ forms of the large subunit and the β subunit of the ( from brain and kidney preparations was examined using rabbit antiserum prepared against the purified holo lamb kidney enzyme. As previously reported by Sweadner ((1979) J. Biol. Chem. 254, 6060–6067) phosphorylation of the large subunit of the ( in the presence of Na+, Mg2+, and [ revealed that dog and, very likely, rat brain contain two forms of the large subunit (designated α and α+) while dog, rat, and lamb kidney contain only one form (α). The cross-reactivity of the α and α+ forms in these preparations was investigated by resolving the subunits by SDS-polyacrylamide gel electrophoresis. The separated polypeptides were transferred to unmodified nitrocellulose paper, and reacted with rabbit anti-lamb kidney serum, followed by detection of the antigen-antibody complex with 125I-labeled protein A and autoradiography. By this method, the α and α+ forms of rat and dog brain, as well as the α form found in kidney, were shown to cross-react. In addition, membranes from human cerebral cortex were shown to contain two immunoreactive bands corresponding to the α and α+ forms of dog brain. In contrast, the brain of the insect Manduca sexta contains only one immunoreactive polypeptide with a molecular weight intermediate to the α and α+ forms of dog brain. The β subunit from lamb, dog and rat kidney and from dog and rat brain cross-reacts with anti-lamb kidney ( serum. The mobility of the β subunit from dog and rat brain on SDS-polyacrylamide electrophoresis gels is greater than the mobility of the β subunit from lamb, rat or dog kidney. 相似文献
7.
A Fourier analysis of the distributions of different types of amino acids in the sequence of tropomyosin shows strong 14th-order peaks in the profiles of both negatively charged and non-polar amino acids, with a period of residues and an overall repeat length of 275 ± 2 amino acids, which is shorter than the sequence length of 284 amino acids. Both peaks are statistically significant and confirm Parry's work (1974, 1975b). The regularities are analysed in terms of an assumed supercoil structure in which two α-helices lie parallel and in register to form a supercoil with a pitch of 137 Å. These molecules are then assumed to overlap end-to-end by eight to nine amino acids so that the periodicity is continuous along an extended filament of linked tropomyosin molecules. The periodic features are stronger in the outer surface of the molecule away from the core of the supercoil. The sequence divides into 14 bands which each have a narrow zone of net positive charge and a broader negatively charged zone. Overlapping every positive zone is a hydrophobic zone which always has at least one non-polar group on the outer surface. Anomalies in the charge distribution are found near the molecular ends and close to Cys190. These are attributed to the end-to-end overlap site and the troponin binding site.In the thin filament the 137 Å pitch supercoil would make seven half-twists relative to the twisted actin helix along a 385 Å length, so that a pair of adjacent bands would be oriented equivalently with respect to a pair of actins 28 Å apart. We therefore suggest that the bands (each containing one zone of each type) should be divided alternately into two series, α and β. Every pair of bands is residues long and each of the seven pairs corresponds with one segment of the 42-residue gene duplication repeat observed previously in the sequence. The disparity between the periods of 42 and is overcome by deletions and insertions. The -residue periodicity is not simply a consequence of the supercoil structure or gene duplication but is probably a result of adaptation to the spatial periodicity of the actin helix in muscle. Although the α and β bands are alike in general, they differ systematically in detail and the α bands are more regular than the β.We propose that the seven α and seven β bands are alternative sets of sites which bind equivalently to complementary groups of sites on seven actins in the “relaxed” and “active” states of muscle, respectively. In each band the negative zone probably attaches to actin by magnesium bridges and the hydrophobic zone by direct contacts with the narrow outer edge of the supercoil. Since the supercoil twists 90 ° relative to actin on passing between adjacent α and β bands, a quarter rotation of the whole tropomyosin molecule would detach one set of seven sites and attach the other, allowing a highly co-operative switch mechanism. 相似文献
8.
S D Dunn 《Biochemical and biophysical research communications》1978,82(2):596-602
ATPase activity was restored to the inactive coupling factor, F1ATPase, of strain AN120 () by reconstitution of the dissociated complex with an excess of wild-type α subunit. Large excesses of α gave the highest levels of activity. The other subunits which are required for the reconstitution of ATPase activity, β and γ, did not complement the mutant enzyme. These results indicate that the α polypeptide of the AN120 ATPase is defective. 相似文献
9.
A complex between EcoRI restriction endonuclease and cognate DNA fragment, 5′-G-A-A-T-T-C C-T-T-A-A-G-5′, has been crystallized. The space group is P4212 with . The unit cell contains four enzyme monomers plus two duplex DNA fragments in an asymmetric unit. High quality crystals of the enzyme alone have also been obtained. 相似文献
10.
The adenosine deaminase of the digestive diverticulum of the bay scallop was purified and electrophoresis of the purified enzyme yielded a single enzymatically active band at several different pH values. A molecular weight of 130,000 was estimated using gel filtration and sucrose density gradient centrifugation. The enzyme had spectral properties typical of simple proteins and its isoelectric point proved to be 4.8. The scallop enzyme was stable at room temperature from pH 5.0 to 7.0, and in this range it was exceptionally resistant to heat inactivation.The effect of the substrate, adenosine, on the reaction velocity was followed over a 10,000-fold concentration range, and no deviation from Michaelis-Menten kinetics was observed. The following rate equation applies to the enzyme: .The effect of pH on the reaction, using adenosine as the substrate, was studied; and it was found that pH had a much greater effect on the α parameter of the rate equation than on the β parameter and that pH had little effect on the apparent activation energy of either parameter. The apparent activation energy of the β parameter was 12.2 kcal with adenosine as the substrate, while the apparent activation energy of the α parameter was zero. The α parameter of the rate equation, using other substrates, was also insensitive to temperature. 相似文献
11.
H. Gehring P. Christen G. Eichele M. Glor J.N. Jansonius A.-S. Reimer J.D.G. Smit C. Thaller 《Journal of molecular biology》1977,115(1):97-101
The mitochondrial isoenzyme of aspartate aminotransferase (E.C. 2.6.1.1) has been isolated from chicken heart in an electrophoretically and immunologically homogeneous form. Large, well-diffracting single crystals of this enzyme, a dimeric molecule with a molecular weight of 90,000, have been grown by vapour phase diffusion against polyethylene glycol solutions. The crystals belong to space group P1. The unit cell, with the dimensions , α = 85.3 °, β = 109.2 °, γ = 115.6 °, contains a single dimer. The diffraction pattern extends to at least 2.1 Å resolution. 相似文献
12.
Nucleotide sequence of the genes coding for alpha, beta and gamma subunits of the proton-translocating ATPase of Escherichia coli 总被引:12,自引:0,他引:12
H Kanazawa T Kayano K Mabuchi M Futai 《Biochemical and biophysical research communications》1981,103(2):604-612
A nucleotide sequence of 2328 base pairs comprising a portion of the gene cluster for the proton-translocating ATPase of was determined. The sequence covers most of the gene for α subunit, the entire gene for γ subunit and the amino terminal portion of the gene for β subunit, along with the flanking regions of these genes. The amino acid sequences of these subunits deduced from the DNA sequences indicate that the α and γ subunits have 513 and 287 amino acid residues, respectively. A possible secondary structure for each subunit was estimated from the inferred primary structure. The intercistronic regions between the genes for α and γ and between γ and β are 49 and 26 base pairs, respectively. The significance of codon usage in these genes is discussed in correlation with their expression. 相似文献
13.
A method to separate phycoerythrin 545, isolated from the cryptomonad alga, , into two subunits has been developed. The method uses no denaturants (urea, guanidine, detergent) but relies on dissociation of the dimeric protein and subsequent aggregation of the β subunit at pH 3.0. The absorption spectra and amino acid composition of the subunits are presented. The spectra of the α subunit was red-shifted relative to β in both pH 3.0 buffer and in acidic 8.0 M urea. 相似文献
14.
《Biochimica et Biophysica Acta (BBA)/General Subjects》1987,925(1):74-82
Under native purification conditions, an oligomeric form (Mr = 230 000) and monomeric form (37 000) of protein B23 were purified by affinity chromatography. Both forms were identified by Western blot immunoassay and ELISA. The molecular weight of the oligomeric form of protein B23 was estimated to be 230 000 with a Stoke's radius and a sedimentation coefficient of 51 Å and 10 S, respectively. The oligomer (230 kDa) of protein B23 was dissociated into monomers (37 kDa) by treatment with 7 M urea. Quantitation of the monomer by gel scanning densitometry indicated that the oligomeric form of protein B23 is a hexamer containing four α and two β monomers (37 kDa). A trace amount of nucleic acids (amounting to less than 3% of the total mass) was detected in the affinity-purified oligomers of protein B23. Protein B23 may be a structural element which is involved in ribosome transport or assembly in the nucleus. 相似文献
15.
16.
M Futai 《Biochemical and biophysical research communications》1977,79(4):1231-1237
The three major subunits (α, β and γ) of the coupling factor, F1 ATPase, of were separated and purified by hydrophobic column chromatography after the enzyme was dissociated by cold inactivation. The ability to hydrolyze ATP was reconstituted by dialyzing the mixture of subunits against 0.05 M Tris-succinate, pH 6.0, containing 2 mM ATP and 2 mM MgCl2. A mixture containing α, β and γ regained ATP hydrolyzing activity. Individual subunits alone or mixtures of any two subunits did not develop ATPase activity, except for a low but significant activity with α plus β. The reconstituted ATPase had a Km of 0.23 mM for ATP and a molecular weight by sucrose gradient density centrifugation of about 280,000. 相似文献
17.
Carlos E. Argaraña Carlos A. Arce Hector S. Barra Ranwel Caputto 《Archives of biochemistry and biophysics》1977,180(2):264-268
In vitro incorporation of [14C]tyrosine into the C-terminal position of the α subunit of tubulin was not affected by 4 mm cycloheximide. This inhibitor of protein synthesis was used for in vivo experiments. The in vivo incorporation of [14C]tyrosine into soluble brain protein of cycloheximide-treated rats was 10% of that of untreated rats. Treatment with vinblastine sulfate of the soluble brain protein showed that the incorporation of [14C]tyrosine into tubulin was higher in cycloheximide-treated than in untreated rats with respect to the incorporation into the total soluble protein. In the case of cycloheximide-treated rats, about 60% of the radioactivity incorporated into protein was released by the action of carboxypeptidase A, whereas 10% was liberated from the protein of untreated rats. The radioactive compound released by the action of carboxypeptidase A was identified as [14C]tyrosine. The α and β subunits of tubulin from animals that received [14C]tyrosine were separated by polyacrylamide gel electrophoresis. The radiosactivity ratio of subunits of tubulin from cycloheximide-treated rats was threefold higher than that of untreated rats. When a mixture of [14C]amino acids was injected, the radioactivity ratio of subunits of tubulin was similar for cycloheximide-treated and untreated rats. The results reported are consistent with the assumption that the α subunit of tubulin can be tyrosinated in vivo. 相似文献
18.
A series of mirror image () forms of prostaglandins F2 and E2 have been compared for potency in a hamster antifertility test. In the PGF2 series, -compounds surveyed had less potency than corresponding natural structures. For the PGE2 series, 11α-(15S)-ent-PGE2 methyl ester was 10-fold more potent than PGE2. Altering the C-9 hydroxy configuration in the PGF2 series from the natural α to β decreased potency dramatically for compounds tested. 相似文献
19.
Robert L. Forti William M. Mitchell Robert J. Workman James T. Forbes Walter C. Hubbard 《Prostaglandins & other lipid mediators》1983,26(3):409-420
Aspirin, indomethacin, and phenbutazone at 50 μM concentration inhibit cyclooxygenase in cultured human foreskin fibroblasts as evidence by the suppression of the major prostaglandin species which aaccumulate in the culture medium. In contrast to data reported for mouse interferon on target mouse cells, these agents have no effect on the introduction of antiviral activity by human α and β interferons. Similarly, these agents have no effect on interferon induced inhibition of cell growth or on interferon induced natural killer cell activity. 相似文献
20.
Two crystal modifications of the monomeric haemoglobin from the flatworm Dicrocoelium dendriticum have been obtained by vapour diffusion against buffered polyethylene glycol solutions. Both the triclinic and hexagonal crystals contain cyanomethaemoglobin. The triclinic modification, space group PI, , with two molecules, Mr = 16,750 each, per unit cell, has been selected for a detailed crystallographic study. 相似文献