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1.
Boronic acid derivatives of good peptide substrates of the serine proteases cause slow-binding inhibition, manifested as biphasic binding (Kettner and Shenvi: J. Biol Chem. 259:15106-15114, 1984). These inhibitors are thought to act as reaction-intermediate analogs. Three peptide boronic acids--Ac-Pro-boro-Val-OH, DNS-Ala-Pro-boro-Val-OH, and Ac-Ala-Ala-Pro-boro-Val-OH--were chosen for far-ultraviolet circular dichroism (CD) studies in order to determine whether the second phase involves a conformational change of pancreatic elastase. The dipeptide is a simple competitive inhibitor (Ki = 0.27 microM) and the latter are slow-binding inhibitors (Ki = 16.4 and 0.25 nM, respectively). Spectral deconvolution and correction for the formation of antiparallel beta-sheet by the peptide inhibitor itself indicate that there is no significant change in the secondary structure of the enzyme in either the initial or final inhibitor complex. A kinetic experiment confirmed that the slow-binding step was not associated with a CD spectral change, and that therefore a protein conformational change was not responsible for the slow binding. 相似文献
2.
Marion T. Hierowski 《FEBS letters》1983,154(1):92-96
Quantitative analyses of LH-RH-like membrane receptors were performed in five tumors from the transplantable Dunning R3372H rat prostatic adenocarcinoma. The binding of D-Trp6-LH-RH, an agonist of LH-RH, was observed in all 5 tumors. The antagonist [Ac-Dp-Cl-Phe1,2,D-Trp3,D-Lys6,D-Ala10]-LH-RH was bound to 4 tumors. The apparent equilibrium dissociation constant (Kd) for D-Trp6-LH-RH receptor was from 2.6–3.9 × 10?10 M. The apparent equilibrium Bmax values (maximum number of binding sites) were from 17.2–86.0 fmol/mg membrane protein for D-Trp6-LH-RH receptor. The Kd for the antagonist was from 2.4–2.7 × 10?10 M and the Bmax values were from 35.5–66.0 fmol/mg membrane protein. Similar binding studies performed in 6 normal rat prostates showed no binding capacities. 相似文献
3.
Bromo[1-14C]acetyl-CoA has been prepared from CoASH and the N-hydroxysuccinimide ester of bromo[1-14C]acetic acid, and unlabeled bromoacetyl-CoA by reaction of CoASH with bromoacetyl bromide. The products were purified by high-pressure liquid chromatography. Purified bromoacetyl-CoA was characterized, and found to be a potent alkylating agent with a substantial stability in aqueous solution: it decomposed at 30 degrees C and pH 6.6 and 8.0 with halftimes of 3.3 and 2.5 h, respectively. The major breakdown products were CoASH and CoAS X CO X CH2 X SCoA. Bromo[1-14C]acetyl-CoA has been used to affinity label the acetyl-CoA binding site of 3-hydroxy-3-methylglutaryl-CoA synthase from ox liver. It was found to irreversibly inhibit the enzyme activity and bind covalently with a stoichiometry for complete inhibition of about 0.8 mol/mol enzyme dimer. 相似文献
4.
Sites of Tubulin Polymerization in PC 12 Cells 总被引:2,自引:0,他引:2
The site at which tubulin enters into polymer in the neuritic process is a very important datum in terms of our understanding of the mechanism of transport of the microtubular cytoskeleton out the axon. If the form of tubulin being transported out the axon is the microtubule, then assembly of tubulin into microtubules should occur at or near the cell body; if, however, the form of tubulin transported is free tubulin dimer, then assembly can occur at any free microtubule end out the neurite. We have injected a fluorescent analog of tubulin into differentiated PC 12 cells and used differential extraction protocols to extract free dimer but not microtubules. We have imaged these cells before and after extraction by low-light-level video fluorescence microscopy and have used image analysis to examine the sites of tubulin incorporation into polymer or other unextracted components as a function of time. We find that tubulin in the distal reaches of the neurite is found initially as monomer and that its appearance in the unextracted component occurs later. This pattern of appearance of fluorescent tubulin initially in the soluble fraction and later in the unextractable component is qualitatively similar to that reported by other workers for biotinylated tubulin, but we see a larger gap between the rates of appearance in soluble fraction and in polymer. Quantitative analysis of fluorescence intensities in the two compartments with distance out the neurite reveals substantial variation between different neurites: In some neurites, the pattern of variation of unextracted/total tubulin suggests that tubulin enters into the unextracted component primarily near the cell body and that this unextracted component moves out the neurite with time, and in other neurites it suggest that monomer adds into microtubule ends staggered out the neurite. In no case do we see a pattern suggesting that distal addition predominates. These analyses of fluorescence intensities in extracted and unextracted neurites suggest that both transport of polymerized microtubules and monomer addition onto staggered microtubule ends occur in PC12 neurites and that in individual neurites one or the other of these two behaviors may predominate. 相似文献
5.
Summary Many plant cell walls are constructed according to a helicoidal pattern that is analog to a cholesteric liquid crystal order. This raises the question whether the wall assembly passes through a true but temporary liquid crystal state. The paper focuses on experiments performed from aqueous suspensions of extracted quince slime, i.e., a cellulose/glucuronoxylan wall composite that presents a helicoidal order when observed in situ, within the enlarged periplasm of the seed epidermal cells. Experiments carried out in acellular conditions showed that a spontaneous reassociation into a helicoidal order can be obtained from totally dispersed suspensions. The ultrastructural aspect of the reassembled mucilage suspension was different according to the resin used (LR White or nanoplast, a water-soluble melamin resin). It was always typically polydomain, and when an order was visible it was cholesteric-like and similar to the in situ native organization. Transition states with many imperfections expressed the difficulty of the system to reassemble in the absence of constraining surfaces. The possible intervention of glucuronoxylan (GX) in the ordered assembly of the microfibrils was checked by: (1) progressive extraction of GX by trifluoroacetic acid (TFA). The extraction was associated to a control of the fraction by analysis of uronic acid contents and observation at the electron microscope level. Extraction of GX provoked the formation of a flocculent mass, the flocculation being more intense when the TFA was more concentrated; (2) progressive change of pH in order to analyze the influence of pH on flocculation. Low pH (ca. pH 3) led also to a flocculation of the suspension, but the floc was reversibly lost after dialysis against distilled water. The results indicate the antifloc role of the GX due to the anionic charges carried by the side-chains. However, the function of GX as helper twisting agent in the cholesteric-like reassembly must not be ruled out. 相似文献
6.
The effects of aluminium (Al), manganese (Mn), zinc (Zn), copper (Cu), boron (B), iron (Fe), gallium (Ga), scandium (Sc) and lanthanum (La) on growth of an Al-tolerant and an Al-sensitive line of wheat (Triticum aestivum L.) were measured in solution culture. The concentrations of nutrients in the basal nutrient solution were (M) 500 Ca, 100 Mg, 300 K, 600 N (150 NH4, 450 NO3), 600 SO4, 2.5 P, 3 B, 2.5 Fe, 0.5 Zn, 0.5 Mn, 0.1 Cu at a pH of 4.7. The major solution nutrient concentrations were maintained at the nominal concentration with monitoring, frequent additions and weekly renewal. Differentiation in yield between the Al-tolerant and Al-sensitive line only occurred in the presence of Al indicating that, in the long term, none of the other metals tested could be used as an analog for Al. The visual symptoms in the roots of Cu toxicity (in both lines) and Al toxicity (in the sensitive line) were similar. The solution concentration (M) at which yield of the roots of the tolerant line was reduced by 50% was, in order of increasing tolerance, Cu 0.5, Sc 1.1, La 7.1, Ga 8.6, Al 15, Zn 19, Fe 84, B 490 and Mn 600. 相似文献
7.
Takeshi Nishimura Setsuko Yamamoto Takaaki Yamamoto Munekiyo Kaneko Youichi Hara 《Prostaglandins & other lipid mediators》1996,51(2):149-159
The antithrombotic effect of topical application of the 3-oxamethano-prostaglandin (PG) I1 analog, SM-10902 in the microcirculation and in vitro antiplatelet functions of its active form SM-10906 were estimated in comparison with PGI2 and PGE1. In rat platelets, SM-10906 evoked accumulation of intracellular cyclic adenosine 3′,5′-monophosphate, and exhibited antiaggregatory and disaggregatory activities, which were all enhanced by the phosphodiesterase inhibitor theophylline. Additionally, SM-10906 was shown to inhibit platelet adhesion to collagen in human platelet-rich plasma. PGI2 and PGE1 also showed in vitro antiplatelet effects in the order of PGI2 > SM-10906 ≥ PGE1. SM-10902 exhibited a dose-dependent antithrombotic effect in the guinea pig mesenteric arteriole by a topical application, and this activity might be exerted by the antiplatelet functions of SM-10906. Although SM-10906, PGI2 and PGE1 also showed the antithrombotic effects, SM-10902 was the most potent. In conclusion, the present studies indicate that an external topical preparation of SM-10902 may be useful for the therapy of peripheral circulatory insufficiency. 相似文献
8.
昆虫卵黄发生研究进展 总被引:19,自引:4,他引:15
昆虫卵黄发生研究进展李乾君,龚和,管致和(中国科学院动物研究所北京100080)(北京农业大学植保系北京100094)昆虫卵的成熟一般分为三个时期--卵黄发生前期(Previtellogenicstage)、卵黄发生期(vitellogenicsta... 相似文献
9.
A complete synthesis of 1-O-hexadecyl-2-O-N-(heptadec-8-cis-enyl)carbamyl-sn-glycero-3-Phosphocholine, a novel analog of phosphatidylcholine, has been described. Each step is simple to perform and gives the desired products in high yield. Also, some of the intermediates formed during the synthesis have been efficiently utilized to prepare 1-O-hexadecyl-2-O-oleyl-sn-glycero-3-phosphocholine, 1-O-hexadecyl-2-oleoyl-sn-glycero-3-phosphochloine and 3-O-hexadecyl-2-oeloyl-sn-glycero-1-phosphocholine. These phosphatidylcholine (PC) analogs are useful for studying the possible role of phospholipases in the capture and lyses of liposomes in vivo. 相似文献
10.
Venkataraman Amarnath Traci L. Miller Arthur D. Broom 《Biochimica et Biophysica Acta (BBA)/General Subjects》1984,800(3):207-213
The synthesis of a high-molecular-weight, putatively all-syn DNA analogue, poly(8-bromo-2′-deoxyadenylic acid), is described. The syn → anti transition was shown to be both salt and temperature dependent. Conditions were found which favored ‘normal’ Watson-Crick pairing and duplex formation with poly(dT). 相似文献