Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P50583 (asymmetrical)
 12,197 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Temperature-induced conformational changes in the anticodon region of yeast tRNATyr were studied by EPR spectroscopy. The spin label 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl was attached to the N6-(delta2-isopentenyl)-adenosine residue in tRNATyr, previously made reactive by iodination. The labelled tRNATyr gave an asymmetrical triplet spectrum typical of rapidly tumbling nitroxide, with a rotational correlation time (tauc) of 0.65 ns. Spin-labelled tRNATyr was exposed to heating and cooling in three different buffers each with or without MgCl2. In each case the Arrhenius plot of --log tauc vs. inverse absolute temperature gave two straight lines, intersecting at a critical temperature (tcr). Above tcr, the anisotropy of the spectrum was not reduced and the activation energy of motion increased, indicating that the transition is associated with a conformational change of the macromolecule. Transitions in 0.05 M potassium phosphate (pH 8.0) and 0.02 M Tris - HC1 (pH 7.0) were observed at potassium phosphate (pH 8.0) and 0.02 M Tris - Hc1 (pH 7.0) were observed at approx. 37 degrees C. When 0.01 M mgCl2 was present in these buffers, transitions were shifted to 46 degrees and 53 degrees C, respectively. Transitions in 0.01 M sodium cacodylate were observed at temperatures which are significantly lower. Since all these transitions occur at temperatures considerably below those required to melt the helical regions of tRNA, and at least approximately 10 degrees C below those reported to break tertiary interactions, it is supposed that they reflect some reorientation of the anticodon region, e.g. a change in tilt of the bases.
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PMID:Conformational changes in yeast tRNATyr revealed by EPR spectra of spin-labelled N6-(delta2-isopentenyl)-adenosine residue. 20 Feb 69

The physicochemical properties of nuclear and cytosolic glucocorticoid-binding components from corticoid-sensitive (CS) and corticoid-resistant (CR) mouse lymphoma P1798 cells have been compared. Nuclei or cytosol fractions were prepared from lymphocytes that had been labeled at 37 or 4 degrees, respectively, with 30 nM [3H]triamcinolone acetonide ([3H]TA). [3H]TA was extracted with 0.6 M KCl, 10 mM spermidine, or 4.5 mM MgCl2 from CS nuclei and with 0.6 M KCl or 10 mM spermidine from CR nuclei. As reported previously, nuclear-associated [3H]TA in CR cells was resistant to extraction with mM concentrations of MgCl2. Loss of bound steroid during extraction with 0.6 M KCl was minimized by including the chymotrypsin inhibitor, carbobenzoxy-L-phenylalanine, in the extraction buffer. The inhibitor was not required during extraction with spermidine or MgCl2. Nuclear and cytosolic extracts were examined by analytical agarose gel filtration and glycerol density gradient centrifugation under high salt (0.6 M KCl) conditions. The glucocorticoid-binding component in KCl, spermidine, and MgCl2 extracts from CS nuclei was considerably larger and more asymmetrical [Stokes radius, 57 to 59 A; sedimentation coefficient, 3.64 to 3.70S; molecular weight, 90,000 daltons; frictional ratio, 1.8; axial ratio (prolate ellipsoid), 15] than the [3H]TA-macromolecular complex in KCl and spermidine extracts from CR nuclei[Stokes radius, 29 A; sedimentation coefficient, 3.23 to 3.30S; molecular weight, 40,000 daltons; frictional ratio, 1.25; axial ratio (prolate ellipsoid), 5]. Control experiments showed that the smaller size of the glucocorticoid-binding component in CR nuclei was probably not due to cleavage of a larger, CS-like complex during the extraction procedure. The larger size of the CS [3H]TA complex did not appear to result from aggregation of s a smaller species. No difference in physicochemical parameters of the binding component was observed if cells were labeled with [3H]dexamethasone instead of [3H]TA. However, [3H]dexamethasone complexes were less stable than those formed with [3H]TA as indicated by considerable dissociation of [3H]dexamethasone during gel filtration and gradient centrifugation. This may be due to the 3- to 5-fold lower relative binding affinity of [3H]dexamethasone. Analysis of [3H]TA-labeled cytosol by gel filtration and gradient centrifugation revealed the presence of a single binding component with physicochemical properties similar to those of nuclear [3H]TA complexes from the same strain of tumor. These results suggest that previously described differences in extractability of nuclear-associated [3H]TA between the CS and CR strains of mouse lymphoma P1798 and the lack of response of CR P1798 to glucocorticoid administration may be due, at least in part, to the presence of an altered glucocorticoid-binding component in the resistant tumor cells.
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PMID:Physicochemical differences between glucocorticoid-binding components from the corticoid-sensitive and -resistant strains of mouse lymphoma P1798. 47 39

The cuticle of the gill lamina of the crayfish Astacus leptodactylus (E), mechanically isolated, was mounted in an Ussing chamber and examined for its electrical properties. The cuticle of the gill lamina obtained from exuviae had similar properties. When perfused with artificial fresh water (AFW) outside and Van Harreveld solution (VH) inside, the transcuticular potential Voi was negative with respect to the inside, and close to the equilibrium potential for Cl- (ECl-). CH3COO-, HCO3-, SO4(2-) and cations (Na+, K+, Ca2+, Mg2+ and NH4+) behaved as impermeant ions with respect to Cl-. A decrease of pH (brought about with CO2) from 8.5 to 6.0 in AFW, VH or both had no effect on the potential. The cuticle area specific conductance was 20-30 mS/cm2 when superfused with AFW outside and VH inside. The conductance decreased linearly with log [Cl-] when Cl- was replaced by CH3COO-. Rectification was obvious when internal Cl- was reduced to 5 mmol/l. The Cl- selectivity of the cuticle could also be demonstrated in perfusing the cuticle with a single salt (NaCl, KCl, CaCl2, MgCl2 or LaCl3) and in diluting that salt on one side of the preparation or in replacing Cl- by CH3COO-, SO4(2-) and HCO3-. The potential changed almost linearly with log [Cl-] and was close to ECl-. The inner face of the cuticle was found to be slightly less selective than the outer face. The relative permeabilities were calculated to be: PCl- = 1, PNa+ = 0.001, PHCO3- = 0.0006, PCH3COO- = 0.0002. The dilution of a Cl- -free salt resulted in a cationic potential. The relative permeabilities of cations (NH4+, K+, Na+, Ca2+ and Mg2+) were found to range within a factor 2. The permeability of the cuticle to HCO3-, CH3COO- and SO4(2-) was 2-5 times lower. The cuticle conductance was linearly related to the activity of the salt perfusing the two sides of the preparation at equal concentrations. The molar area specific conductance to chloride salts was 14 (mS/cm2)/(mmol/l). That of Cl- -free salts ranged from 1 to 20 (microS/cm2)/(mmol/l) depending on the salt used. It was deduced that PCl- is 2 X 10(-3) cm/s and that all the other ions tested have permeabilities of 10(-7)-10(-6) cm/s. With large intensity current pulses the cuticle exhibited rectifying properties and an asymmetrical behaviour. Increasing the pH of the perfusing solution reduced the transcuticular potential established with a Cl- gradient.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Ionic permeabilities of the gill lamina cuticle of the crayfish, Astacus leptodactylus (E). 241 Jun 7

The DNA-binding form of the calf uterine androgen receptor (AR) was subjected to limited protease digestion using chymotrypsin, trypsin and a rat prostate cytosol protease. The properties of the generated polypeptide fragments were identified and compared with those of the intact AR. Physicochemical characterization was achieved through sedimentation analysis, gel filtration chromatography and DEAE anion exchange chromatography. Intactness of functional binding domains was evaluated by measuring the retention of steroid- and DNA-binding capacity. Under non-denaturing conditions the intact AR is a highly asymmetrical molecule with a Stokes radius (RS) of 45A, a sedimentation coefficient of 4.3S and a relative molecular mass of 80,000 daltons. This form of AR has an intrinsic binding affinity for DNA and was eluted from DNA-cellulose with 9 mM MgCl2. Chymotrypsin produced a more globular polypeptide (RS: 31A; 3.1S; 41,000 daltons) with a decreased net negative charge. This fragment also displayed DNA-binding affinity but required a higher concentration of MgCl2 (14 mM) for DNA-cellulose elution, indicating an increased affinity for DNA. The observed reduction in molecular size upon chymotrypsin treatment was confirmed when analysed by SDS-polyacrylamide gel electrophoresis after covalently labelling of the AR with [3H]R1881. Rat prostate cytosol contains a protease which is very active in generating an AR polypeptide with an increased affinity for DNA, without changing the AR net negative charge (RS: 33A; 3.7S; 51,000 daltons). The specificity of this protease remained unknown since none of a large number of inhibitors was able to inactivate this enzyme. The fragment generated is different from that obtained with chymotrypsin since significant differences in size as well as in charge were measured. Trypsin treatment generated a much smaller polypeptide (RS: 25A; 2.9S; 30,000 daltons) which had lost its DNA-binding capacity, but not its steroid binding site. This form probably represents the so-called meroreceptor. When intact AR was treated sequentially with prostate cytosol and trypsin, a polypeptide fragment with identical properties was obtained, indicating the spatial separation of two of the proteolytic cleavage sites. These studies provide evidence for the distinct nature of the molecular domains for androgen and DNA interaction on the calf uterine AR.
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PMID:Analysis of steroid- and DNA-binding domains of the calf uterine androgen receptor by limited proteolysis. 330 38

In freeze-fracturing human erythrocytes beside the intramembraneous particles fibrillar structures can be observed. They are in close relation to the intramembraneous particles and occur frequently under special conditions. 1. In erythrocytes incubated with MgCl2-solutions (25-100 mM) striking fibrillar structures are mostly located on the E-fracture face or extend from the E-face to the cytoplasmic surface aspect of the membrane. Fibrillar structures located on the P-face or extending from the P-face to the extracellular surface aspect do not often occur. This asymmetrical distribution indicates that elements pulled out from the cytoplasmic membrane half play a special role in the fibril formation. Intermediate forms between particles and fibrillar structures can occur. 2. By use of the microtome technique for freeze-fracturing in a frozen 60 vol.-% glycerol solution at temperatures upward from -100 degrees C melting can occur at the knife edge during cutting. In connection with the formation of a melting zone in isolated erythrocyte membranes incubated in 60 vol.-% aqueous glycerol a somewhat modified (unfinished) membrane splitting is possible. Lying side by side both the membrane fracture halves remain connected by the unsplit part of the membrane. In the region where they are connected fibrils between the two fracture faces occur indicating a deformation process during the particle formation. The results give further evidence that plastic deformation during freeze-fracturing takes place in the formation of the intramembraneous particles in native erythrocyte membranes.
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PMID:Variations in the appearance of membrane particles after various pretreatments. 678 61

The diadenosine 5',5'''-P1,P4-tetraphosphate (asymmetrical) hydrolase (EC 3.6.1.17) from human placenta has been purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephacel, gel filtration on Sephadex G-100, and affinity elution from red Sepharose. The enzyme is a single polypeptide of M(r) 19,200. It exhibits maximum (100%) activity at pH 7.3 in the presence of 3 mM MgCl2 and 60, 50, and 40% of the activity in 1 mM CoCl2, 0.1 mM ZnCl2, and 0.5 mM MnCl2, respectively. The Km value calculated for diadenosine tetraphosphate in the presence of Mg2+ is 10 microM and in the presence of Zn2+ 40 microM. Adenosine 5'-tetraphosphate, guanosine 5'-tetraphosphate, and fluoride proved to be inhibitors of the diadenosine tetraphosphate hydrolase; the I50 values were 6, 10, and 20 microM, respectively. Diguanosine tetraphosphate, bis-2,6-diaminopurine beta-D-ribofuranoside tetraphosphate, and diadenosine pentaphosphate were substrates for the hydrolase; relative velocities of hydrolysis estimated for 0.5 mM diadenosine tetraphosphate and these other substrates were 1:0.51:0.44:0.20, respectively. Diadenosine tetraphosphate analogues with P2-P3 bridges such as -CF2-, -CCl2-, and -CH2- were hydrolyzed to adenosine 5'-phosphate and the corresponding adenosine 5'-triphosphate analogue.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Human placental (Asymmetrical) diadenosine 5',5'''-P1,P4-tetraphosphate hydrolase: purification to homogeneity and some properties. 838 Oct 42