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排序方式: 共有232条查询结果,搜索用时 15 毫秒
1.
M Chorev E Roubini R L McKee S W Gibbons J E Reagan M E Goldman M P Caulfield M Rosenblatt 《Peptides》1991,12(1):57-62
Lysine occupies position 13 in the parathyroid hormone (PTH) antagonist, [Nle8,18,Tyr34]bPTH(7-34)NH2. Acylation of the epsilon-amino group in lysine 13 by a hydrophobic moiety is well tolerated in terms of bioactivity: the analog [Nle8,18, D-Trp12,Lys 13 (epsilon-3-phenylpropanoyl),Tyr34]bPTH(7-34)NH2 is equivalent to the parent peptide in its affinity for PTH receptors and its ability to inhibit PTH-stimulated adenylate cyclase in both kidney- and bone-based assays. Truncation of this peptide by deletion of phenylalanyl7 with concomitant removal of the amino-terminal alpha-amino group yielded the analog desamino[Nle8,18,D-Trp12,Lys13 (epsilon-3-phenylpropanoyl),Tyr34]bPTH(8-34)NH2, an antagonist of high potency in vitro (Kb = 4 and 9 nM, Ki = 73 and 3.5 nM in kidney- and bone-based assays, respectively). Also this analog is potentially stable to aminopeptidases present in many biological systems. 相似文献
2.
Frank K. Klemens John C. Severns Ramasamy Tamilarasan David R. McMillin 《Inorganica chimica acta》1996,250(1-2):75-79
This study dealt with the demetalation and remetalation chemistry of the copper-containing protein ceruloplasmin. For the enzyme from human plasma, dialysis against cyanide at 4°C readily removed copper. Although the apoprotein took up copper(I) at the same temperature, the characteristic blue color of the native protein did not return to any significant extent. However, excellent reconstitution occurred when we added copper(I) at room temperature. With porcine ceruloplasmin, it was more practical to carry out the copper removal step at room temperature, but the reconstitution went smoothly at 4°C. With either source of ceruloplasmin, the binding of the six essential copper ions was generally a highly cooperative process, but the results were different when we combined the apoprotein with Hg(II). After the protein took up 2 mercury ions, it would take up only 1–2 more metal ions even after exposure to a large excess of copper(I). In order to accomodate the various experimental results, we have proposed that a reversible conformational change must occur during the demetalation and remetalation processes. During the remetalation process, it is therefore important that metal uptake occurs in the proper sequence. 相似文献
3.
The N-1-Naphthylphthalamic Acid-Binding Protein Is an Integral Membrane Protein 总被引:6,自引:3,他引:3 下载免费PDF全文
N-1-Naphthylphthalmic acid (NPA)-binding protein is a plasmalemma (PM) protein involved in the control of cellular auxin efflux. We re-evaluated the spatial relationship of this protein with the PM of zucchini (Cucurbita pepo L.) hypocotyls. First, Triton X-114 partitioning indicated that the NPA-binding protein was more hydrophobic than most PM proteins. Second, the NPA-binding activity was found to be resistant to proteolytic digestion in membranes. Maximum concentrations of binding sites for NPA were virtually identical in untreated and proteinase K-treated PMs: 19.2 and 20.6 pmol [3H]NPA bound/mg protein, respectively. The insensitivity of the NPA-binding protein was not due to its presence inside tightly sealed vesicles or due to lack of protease activity in the conditions tested. This protein could be made sensitive to proteolytic degradation upon solubilization by 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate in the presence of sodium molybdate. Proteinase K treatment decreased the concentration of binding sites to 0.84 pmol [3H]NPA bound/mg protein from 9.2 for untreated, solubilized PM. Third, this activity could not be solubilized by chaotropic agents or sodium carbonate treatment of intact PM. This study indicates that the NPA-binding protein may be an integral membrane protein and contradicts previously reported findings that suggested that this protein was peripheral to the PM. 相似文献
4.
The type 1 copper in Pseudomonas aeruginosa azurin was studied by electron paramagnetic resonance (EPR) spectroscopy at low microwave frequencies. Partially resolved ligand hyperfine structure was observed in the perpendicular region of the spectra at both S-band (2.4 GHz) and L-band (1.1 GHz). A trial and error method, requiring several hundred simulations, has been used to simulate the low frequency EPR data and yield an optimum value of 30 MHz for ACUx, more than one half that previously reported. The fit between the simulated and experimental data is sensitive to changes in the Euler angles and, in particular, to the angle alpha which rotates the Cu A-tensor about the z-axis. Thus, the A- and g-tensors for copper in P. aeruginosa azurin do not appear to be coincident. A value for the Euler angle beta of at least 10 degrees does not disturb the fit between the simulated and experimental data. These studies demonstrate the advantage of evaluating EPR parameters from simulations at more than one frequency, especially at low frequencies where ligand superhyperfine structure may be resolved for type 1 copper. 相似文献
5.
We have refined a useful incubation method for preparing adducts of tree laccase with inhibitor anions; however, the technique should have wider application. The procedure involves incubating a previously frozen aqueous solution at subzero temperature. Factors influencing the amount of adduct that forms include the nature of the buffer, pH changes that occur at subzero temperatures and the presence of glycerol. In the absence of a glassing agent like glycerol, the phase separation and solute pooling that occur with ice formation help drive adduct formation. The results reveal several important factors to consider in designing or interpreting low-temperature spectroscopic investigations. 相似文献
6.
Low-density lipoproteins (hLDL) and beta-migrating-very-low-density lipoproteins (beta-VLDL) were isolated from the plasma of cholesterol-fed White Carneau (WC) pigeons and low-density lipoproteins (nLDL) were isolated from the plasma of grain-fed WC pigeons. The lipoproteins were radiolabeled with 125I or 131I and injected into normocholesterolemic or hypercholesterolemic WC pigeons to determine their rate of clearance from the plasma. The fractional catabolic rate (FCR) of nLDL and hLDL in normocholesterolemic pigeons averaged 0.202 and 0.206 pools/h.respectively. beta-VLDL was cleared at a significantly slower rate of 0.155 pools/h. The FCR of the same lipoproteins injected into hypercholesterolemic pigeons was reduced by 17% for nLDL, 50% for hLDL and 57% for beta-VLDL, indicating that the effect of hypercholesterolemia on clearance in vivo was different for the three lipoproteins. The FCR of reductively methylated pigeon LDL (MeLDL), which gives a measure of receptor-independent clearance of LDL, was shown previously to be 0.037 pools/h. These studies suggest therefore that LDL and beta-VLDL are cleared from the plasma of normocholesterolemic and hypercholesterolemic pigeons at a rate substantially greater than that predicted for non-specific processes. Despite the reduction in the clearance rate of hLDL and beta-VLDL due to cholesterol feeding, the absolute amount of cholesterol that was cleared from the plasma by these lipoproteins was increased from approx. 200 mg/kg body weight per day in the normocholesterolemic pigeons to greater than 1000 mg/kg body weight per day in the hypercholesterolemic pigeons. This is due principally to the enrichment in cholesterol relative to protein of the lipoproteins isolated from cholesterol-fed pigeons and the failure of hypercholesterolemia to completely inhibit receptor-dependent clearance of LDL and beta-VLDL. The lower rate of clearance of beta-VLDL relative to LDL is in marked contrast to mammalian beta-VLDL, which is cleared much faster than LDL, but is consistent with the lack of apo E on pigeon lipoproteins. Apo E is the apoprotein that is thought to be responsible for the rapid clearance of beta-VLDL in normocholesterolemic mammals. The low rate of beta-VLDL clearance in pigeons also suggests that pigeons lack an apolipoprotein that function like mammalian apo E. 相似文献
7.
Modifications of position 12 in parathyroid hormone and parathyroid hormone related protein: toward the design of highly potent antagonists 总被引:1,自引:0,他引:1
M Chorev M E Goldman R L McKee E Roubini J J Levy C T Gay J E Reagan J E Fisher L H Caporale E E Golub 《Biochemistry》1990,29(6):1580-1586
Truncated N-terminal fragments of parathyroid hormone (PTH), [Tyr34]bovine PTH(7-34)NH2, and parathyroid hormone related protein (PTHrP), PTHrP(7-34)NH2, inhibit [Nle8,18,[125I]iodo-Tyr34]-bPTH(1-34)NH2 binding and PTH-stimulated adenylate cyclase in bone and kidney assays. However, the receptor interactions of these peptides are 2-3 orders of magnitude weaker than those of their agonist counterparts. To produce an antagonist with increased receptor-binding affinity but lacking agonist-like properties, structure-function studies were undertaken. Glycine at position 12 (present in all homologues of PTH and in PTHrP), which is predicted in both hormones to participate in a beta-turn, was examined by substituting conformational reporters, such as D- or L-Ala, Pro, and alpha-aminoisobutyric acid (Aib), in both agonist and antagonist analogues. Except for N-substituted amino acids, which substantially diminished potency, substitutions were well tolerated, indicating that this site can accept a wide latitude of modifications. To augment receptor avidity, hydrophobic residues compatible with helical secondary structure were introduced. Incorporation of the nonnatural amino acids D-Trp, D-alpha-naphthylalanine (D-alpha-Nal), or D-beta-Nal into either [Tyr34]bPTH(7-34)NH2 or [Nle8,18,Tyr34]bPTH(7-34)NH2 resulted in antagonists that were about 10-fold more active than their respective 7-34 parent compound. Similarly, [D-Trp12]PTHrP(7-34)NH2 was 6 times more potent than the unsubstituted peptide but retained partial agonistic properties, although markedly reduced, similar to PTHrP(7-34)NH2. The antagonistic potentiating effect was configurationally specific.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
8.
Persistence of human rhinovirus infectivity under diverse environmental conditions. 总被引:1,自引:1,他引:0 下载免费PDF全文
The persistence of human rhinovirus type 2 and type 14 infectivity was studied under various laboratory conditions designed to mimic those commonly found in the environment. The effects of temperature, ionic strength, protein content, and evaporation were compared. Both viruses were stable (less than 0.3-log decrease in titer) at 6 and 23 degrees c for 24 h in the liquid state regardless of salt or protein additives; a titer decrease of less than 1.0 log was noted at 37 degrees C. However, evaporation at 37 degrees C reduced virus infectivity by 3.2 to 4.5 logs in buffered water, an effect which could be significantly lessened by the addition of bovine serum albumin in saline (2.0- to 2.9-log decrease in titer). These studies support and extend observations by others that the human rhinoviruses retain sufficient infectivity after drying on hard surfaces to permit their transmission to susceptible persons upon contact. 相似文献
9.
E I Solomon R Wang D R McMillin H B Gray 《Biochemical and biophysical research communications》1976,69(4):1039-1042
Magnetic susceptibility measurements on cobalt(II) stellacyanin () have been performed between 2.2 and 50 K. The effective magnetic moment of Co(II) in the protein is 3.91 ± 0.12 (μB). Nonlinear behavior below 3 K evidences the presence of zero-field splitting attributable to a low-symmetry component of the ligand field. The results are consistent with a structural model based on a distorted tetrahedral Co(II) site involving one or more extremely covalent metal-ligand bonds. 相似文献
10.
M Chorev E Roubini M E Goldman R L McKee S W Gibbons J E Reagan M P Caulfield M Rosenblatt 《International journal of peptide and protein research》1990,36(5):465-470
Position 18 in a parathyroid hormone (PTH) antagonist, [Nle8,18,Tyr34]bPTH(7-34)NH2 (ii), was shown to tolerate substitutions by a range of amino acids with retention of inhibitory activity. The effects of hydrophobic substitutions at this position as a means of enhancing binding interactions with the receptor were evaluated. Substitution of Nle at position 18 with either D-Ala, D-Trp, or L-Trp in analog ii or with Trp (D or L) in the recently reported, highly potent antagonist, [Nle8,18,D-Trp12,Tyr34]bPTH(7-34)NH2 (in vitro activities; Kb = 15 nM and Ki = 125 nM), was performed. In terms of activity on renal receptors, one antagonist, [Nle8,D-Trp12,18,Tyr34]bPTH(7-34)NH2, is the most active in vitro PTH antagonist yet reported (Kb = 4 nM; Ki = 30 nM). The rationale for design of this antagonist and the conclusions regarding PTH-receptor interactions are discussed. 相似文献