共查询到20条相似文献,搜索用时 15 毫秒
1.
Lorne J. Brandes Gary M. Queen Frank S. LaBella 《Journal of cellular biochemistry》1998,69(3):233-243
Histamine and polyamines have been implicated in the mediation of cell proliferation. Our previous work linked the growth-modulatory effects of histamine with its binding to intracellular sites in microsomes and nuclei of various tissues. In this study, we identify cytochrome P450 enzymes as a major component of microsomal intracellular sites in hepatocytes and demonstrate that polyamines compete with high affinity for histamine binding to them. Spectral measurement of histamine binding to P450 in liver microsomes resolved high and intermediate affinity binding sites (Ks1 = 2.4 ± 1.6 μM; Ks2 = 90 ± 17 μM) that corresponded to microsomal binding sites (Kd1 = 1.0 ± 0.9 μM; Kd2 = 57 ± 13 μM) resolved by 3H-histamine binding; additional low affinity (Kd3 ∼ 3 mM), and probably physiologically irrelevant, sites were resolved only by 3H-histamine radioligand studies. As determined spectrally, treatment of microsomes with NADPH/carbon monoxide decreased histamine binding to P450 by about 90% and, as determined by 3H-histamine binding, abolished the high affinity sites and reduced by 85% the number of intermediate sites. Spermine competed potently for 3H-histamine binding: in microsomes, Ki = 9.8 ± 5.8 μM; in nuclei, Ki = 13.7 ± 3.1 μM; in chromatin, Ki = 46 ± 33 nM. Polyamines inhibited the P450/histamine absorbance complex with the rank order of potency: spermine > spermidine ≫ putrescine. In contrast, histamine did not compete for 3H- spermidine binding in nuclei or microsomes, suggesting that polyamines modulate histamine binding allosterically. We propose that certain P450 isozymes that modulate gene function by controlling the level of oxygenated lipids, represent at least one common intracellular target of growth-regulatory endogenous bioamines and, as shown previously, of exogenous growth-modulatory drugs including antiestrogens, antiandrogens, and certain antidepressants and antihistamines. J. Cell. Biochem. 69:233–243, 1998. © 1998 Wiley-Liss, Inc. 相似文献
2.
Peter J.M. Van Haastert 《Biochemical and biophysical research communications》1984,124(2):597-604
D. discoideum contains kinetically distinguishable cell surface cAMP binding sites. One class, S, is slowly dissociating and has high affinity for cAMP (Kd = 15 nM, ). A second class is fast dissociating ( about 1 s) and is composed of high affinity binding sites H (Kd ≈ 60 nM), and low affinity binding sites L (Kd = ≈ 450 nM) which interconvert during the binding reaction. Guanine nucleotides affect these three binding types in membranes prepared by shearing D.discoideum cells through Nucleopore filters. The affinity of S for cAMP is reduced by guanine nucleotides from 13 nM to 25 nM, and the number of S-sites is reduced about 50%. The number of fast dissociating sites is not altered by guanine nucleotides, but these sites are mainly in the low affinity state. Half-maximal effects are obtained at about 1 μM GTP, 2 μM GDP and 10 μM Gpp(NH)p(guanyl-5′-yl-imidodiphosphate); ATP and ADP are without effect up to 1 mM. These results indicate that D.discoideum cells have a functionally active guanine nucleotide binding protein involved in the transduction of extracellular cAMP signals via cell surface cAMP receptors. 相似文献
3.
The actions of a number of sodium channel-specific neurotoxins on the uptake of Na-22 by osmotically sensitive membrane preparations from rat brain were studied using a glass-fiber filter assay. Under control conditions, there was Na-22 uptake that reached saturation within 5 min, and was insensitive to tetradotoxin (10 μM). Batrachotoxin (Kdapp = 0.2 μM), veratridine (Kdapp = 1 μM) and grayanotoxin I (Kdapp = 30 μM), which increase sodium conductance in electrogenic membranes, stimulated Na-22 uptake approximately 2-fold over control levels. This additional Na-22 uptake was markedly dependent on the ionic strength of the media, associated with subfractions of the preparation enriched in plasma membranes, and completely inhibited by tetrodotoxin (10 μM). It was highly labile, showing only a minor decrease in activity within the first 4–6 h after preparation of the membranes, but disappearing within 24 h at 4° C. It is suggested that the toxins-activated Na-22 uptake, which is tetrodotoxin-sensitive, results from the actions of these toxins on the macromolecular channel complex which controls resting and action potential sodium conductance. 相似文献
4.
Leon Milewich Terry S. Hendricks Bettie Sue Masters Rene A. Frenkel Paul C. Mac Donald 《Archives of biochemistry and biophysics》1981,211(2):530-536
The estrogen synthetase present in human placental microsomes appears to be dependent on the cooperative interaction of the reduced cofactors NADPH and NADH for optimal activity. Using steady-state concentrations of either cofactor, it was found that while the estrogen synthetase activity followed hyperbolic saturation kinetics with NADPH (Kmapp = 14 μM), the enzyme followed sigmoidal saturation kinetics when the cofactor was NADH, with the half-maximum velocity attained at a cofactor concentration of 1.1 mm. The maximum velocity obtained with NADPH as the cofactor was greater than with corresponding concentrations of NADH. Estrogen synthetase activity in the presence of NADH was not due to NADPH contamination. NADH, in the presence of small concentrations of NADPH (0.5 to 5 μm), stimulated significantly the rate of estrogen formation from androstenedione by placental microsomes and, in addition, the enzyme saturation kinetics changed from sigmoidal to hyperbolic, thus mimicking the effect of NADPH. Estrogen synthetase activity, measured in the presence of 1 mm NADH, was stimulated in a dose-dependent manner by NADPH (Kmapp = 0.4 μM NADPH) and, when the enzyme was measured in the presence of 5 μm NADPH, the activity was stimulated in a dose-dependent manner by NADH (Kmapp = 45 μM NADH). Estrogen synthetase activity measured in the presence of NADH, without and with NADPH (1 μm) remained linear both with time of incubation for approximately 15 min and with microsomal protein concentration up to 3 mg/ml. The apparent Km of estrogen synthetase for androstenedione, when measured in the presence of NADH, was 1 μm. The synergistic interaction between NADH and NADPH in stimulating placental estrogen synthetase activity observed in vitro may, conceivably, take place in vivo in the intact placenta. 相似文献
5.
Robert L. Barbieri Rapin Osathanondh Jacob A. Canick Robert J. Stillman Kenneth J. Ryan 《Steroids》1980,35(3):251-263
The effects of danazol on steroidogenesis in the 16–20 week old human fetal adrenal were examined by studying: 1) danazol binding to adrenal microsomal and mitochondrial cytochrome P-450, and 2) enzyme kinetics of danazol inhibition of the adrenal microsomal 21-hydroxylase and the mitochondrial llβ-hydroxylase. The addition of danazol to preparations of adrenal microsomes or mitochondria elicited a type I cytochrome P-450 binding spectrum. Danazol bound to microsomal cytochrome P-450 with a high affinity apparent spectral dissociation constant (Kg) of 1 μM and with a lower affinity K's of 10 μM. Danazol bound to mitochondrial cytochrome P-450 with a Kg of 5 μM. In addition, danazol competitively inhibited the microsomal 21-hydroxylase (apparent enzymatic inhibition constant KI = 0.8 μM) and the mitochondrial 11β-hydroxylase (KI = 3 μM). These findings demonstrate that low concentrations of danazol directly inhibit steroidogenesis in the human fetal adrenal . 相似文献
6.
Pradeep S. Pallan Chunxue Wang Li Lei Francis K. Yoshimoto Richard J. Auchus Michael R. Waterman F. Peter Guengerich Martin Egli 《The Journal of biological chemistry》2015,290(21):13128-13143
Cytochrome P450 (P450) 21A2 is the major steroid 21-hydroxylase, and deficiency of this enzyme is involved in ∼95% of cases of human congenital adrenal hyperplasia, a disorder of adrenal steroidogenesis. A structure of the bovine enzyme that we published previously (Zhao, B., Lei, L., Kagawa, N., Sundaramoorthy, M., Banerjee, S., Nagy, L. D., Guengerich, F. P., and Waterman, M. R. (2012) Three-dimensional structure of steroid 21-hydroxylase (cytochrome P450 21A2) with two substrates reveals locations of disease-associated variants. J. Biol. Chem. 287, 10613–10622), containing two molecules of the substrate 17α-hydroxyprogesterone, has been used as a template for understanding genetic deficiencies. We have now obtained a crystal structure of human P450 21A2 in complex with progesterone, a substrate in adrenal 21-hydroxylation. Substrate binding and release were fast for human P450 21A2 with both substrates, and pre-steady-state kinetics showed a partial burst but only with progesterone as substrate and not 17α-hydroxyprogesterone. High intermolecular non-competitive kinetic deuterium isotope effects on both kcat and kcat/Km, from 5 to 11, were observed with both substrates, indicative of rate-limiting C–H bond cleavage and suggesting that the juxtaposition of the C21 carbon in the active site is critical for efficient oxidation. The estimated rate of binding of the substrate progesterone (kon 2.4 × 107
m−1 s−1) is only ∼2-fold greater than the catalytic efficiency (kcat/Km = 1.3 × 107
m−1 s−1) with this substrate, suggesting that the rate of substrate binding may also be partially rate-limiting. The structure of the human P450 21A2-substrate complex provides direct insight into mechanistic effects of genetic variants. 相似文献
7.
Philippe Lemaire David R. Livingstone 《Journal of biochemical and molecular toxicology》1994,9(2):87-95
Inhibitors of mammalian cytochrome P450 and P450 reductase were used to investigate the enzymes in flounder (Platichthys flesus) hepatic microsomes involved in the stimulation of NAD(P)H-dependent iron/EDTA-mediated 2-keto-4-methiolbutyric acid (KMBA) oxidation (hydroxyl radical production) by the redox cycling compounds menadione and nitrofurantoin. Inhibitors were first tested for their effects on flounder microsomal P450 and flavoprotein reductase activities. Ellipticine gave type II difference binding spectra (app. Ks 5.36 μM; ΔA max 0.16 nmol-1 P450) and markedly inhibited NADPH-cytochrome c reductase, NADPH-cytochrome P450 reductase, and monooxygenase (benzo[a]pyrene metabolism) activities. 3-aminopyridine adenine dinucleotide phosphate (AADP; competitive inhibitor of P450 reductase) inhibited NADPH-cytochrome c but not NADH-cytochrome c or NADH-ferricyanide reductase activities. Alkaline phosphatase (inhibitor of rabbit P450 reductase) stimulated NADPH-cytochrome c reductase activity seven fold but had less effect on NADH-reductase activities. AADP inhibited nitrofurantoin- and menadione-stimulated KMBA oxidation by 45 and 17%, respectively, indicating the involvement of P450 reductase at least in the former. In contrast, ellipticine had relatively little effect, possibly because, unlike cytochrome c, the smaller xenobiotic molecules can access the hydrophilic binding site of P450 reductase. Alkaline phosphatase stimulated NAD(P)H-dependent basal and xenobiotic-stimulated KMBA oxidation, showing general consistency with the results for reductase activities. Overall, the studies indicate both similarities (ellipticine, AADP) and differences (alkaline phosphatase) between the flounder and rat hepatic microsomal enzyme systems. 相似文献
8.
《Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology》1988,952(2):173-177
The single glutathione S-transferase (EC 2.5.1.18) present in rat erythrocytes was purified to apparent homogeneity by affinity chromatography on glutathione-Sepharose and hydroxyapatite chromatography. Approx. 1.86 mg enzyme is found in 100 ml packed erythrocytes and accounts for about 0.01% of total soluble protein. The native enzyme (Mr 48 000) displays a pI of 5.9 and appears to possess a homodimeric structure with a subunit of Mr 23 500. Enzyme activities with ethacrynic acid and cumene hydroperoxide were 24 and 3%, respectively, of that with 1-chloro-2,4-dinitrobenzene. The Km values for 1-chloro-2,4-dinitrobenzene and glutathione were 1.0 and 0.142 mM, respectively. The concentrations of certain compounds required to produce 50% inhibition (I50) were as follows: 12 μM bromosulphophthalein, 34 μM S-hexylglutathione, 339 μM oxidized glutathione and 1.5 mM cholate. Bromosulphophthalein was a noncompetitive inhibitor with respect to 1-chloro-2,4-dinitrobenzene (Ki = 8 μM) and glutathione (Kis = 4 μM; Kii = 11.5 μM) while S-hexylglutathione was competitive with glutathione (Ki = 5 μM). 相似文献
9.
Some pharmacological properties of ellipticine (E) and its derivatives linked to their interaction with cytochrome P-450 have been investigated with human liver microsomes. 9-Hydroxyellipticine (9-OHE) interacts with human liver cytochrome P-450 exhibiting a type II spectrum (λmax: 428 nm, Ks = 1.1 μM). After incubation with human liver microsomes the E was converted to 9-OHE; 7-hydroxyellipticine was not produced. The cytotoxic effect of this biotransformation has been evaluated on leukemic L1210 cells, in vitro, and found to be equal to those elicited by liver microsomes of control or phenobarbital (PB) pretreated rats. Moreover, 9-OHE and 9-fluoroellipticine (9-FE) strongly inhibit the benzo[a]pyrene hydroxylase (AHH) activity of human liver microsomes (I50 = 2.6 μM and 1.6 μM, respectively) as well as the mutagenesis induced by the polycyclic aromatic hydrocarbon 2-acetylaminofluorene (AAF); 1 μg/plate of each of these compounds is able to inhibit by more than 50% the mutagenicity of 5 μg/plate AAF. 相似文献
10.
Starvation Alters the Apparent Half-Saturation Constant for Methane in the Type II Methanotroph Methylocystis Strain LR1
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When cells of a type II methanotrophic bacterium (Methylocystis strain LR1) were starved of methane, both the Km(app) and the Vmax(app) for methane decreased. The specific affinity (aos) remained nearly constant. Therefore, the decreased Km(app) in starved cells was probably not an adjustment to better utilize low-methane concentrations. 相似文献
11.
Two soluble hexokinases and a particulate hexokinase have been separated and partially purified from spinach leaves. One of the soluble hexokinases showed a high affinity for glucose (Km = 63 μM) which was far greater than that for fructose (Km = 9.1 mM). However, with saturating fructose the activity was twice that with saturating glucose. The particulate hexokinase showed kinetic properties similar to those of this soluble hexokinase. The second soluble hexokinase was distinct in that it was much more active with fructose than with glucose at all concentrations tested, although the Km values for these hexoses (210 μM and 71 μM respectively) were similar. The activity of this hexokinase was stimulated by the monovalent cations K+ and NH4+. 相似文献
12.
《BBA》1986,849(1):121-130
The binding of 3′-O-(1-naphthoyl)adenosinetriphosphate (1-naphthoyl-ATP), ATP and ADP to TF1 and to the isolated α and β subunits was investigated by measuring changes of intrinsic protein fluorescence and of fluorescence anisotropy of 1-naphthoyl-ATP upon binding. The following results were obtained. (1) The isolated α and β subunits bind 1 mol 1-naphthoyl-ATP with a dissociation constant (KD(1-naphthoyl-ATP)) of 4.6 μM and 1.9 μM, respectively. (2) The KD(ATP) for α and β subunits is 8 μM and 11 μM, respectively. (3) The KD(ADP) for α and β subunits is 38 μM μM and 7 μM, respectively. (4) TF1 binds 2 mol 1-naphthoyl-ATP per mol enzyme with KD = 170 nM. (5) The rate constant for 1-naphthoyl-ATP binding to α and β subunit is more than 5 · 104 M−1s−1. (6) The rate constant for 1-naphthoyl-ATP binding to TF1 is 6.6 · 103 M−1 · s−1 (monophasic reaction); the rate constant for its dissociation in the presence of ATP is biphasic with a fast first phase (kA−1 = 3 · 10−3s−1) and a slower second phase (kA−2 < 0.2 · 10−3s−1). From the appearance of a second peak in the fluorescence emission spectrum of 1-naphthoyl-ATP upon binding it is concluded that the binding sites in TF1 are located in an environment more hydrophobic than the binding sites on isolated α and β subunits. The differences in kinetic and thermodynamic parameters for ligand binding to isolated versus integrated α and β subunits, respectively, are explained by interactions between these subunits in the enzyme complex. 相似文献
13.
Intracellular products of peptide transport were analyzed after thin layer chromatography separation by a fluorescamine labeling procedure. Dipeptides penetrated Pseudomonas aeruginosa via 2 transport systems, one of high affinity with KT values from 1–2 μM, and another one of low affinity (KT about 30 μM). A single transport system assumed tripeptide uptake with KT varying from 10–30 μM, depending on the tripeptide used. Peptides entering the cells were rapidly hydrolyzed into their constitutive amino acids. 相似文献
14.
《FEBS letters》1997,407(1):69-72
Hydroxyquinol 1,2-dioxygenase, an intradiol dioxygenase, which catalyzes the cleaving of the aromatic ring of hydroxyquinol, a key intermediate of 2,4-D and 2,4,5-T degradation, was purified from Nocardioides simplex 3E cells grown on 2,4-D as the sole carbon source. This enzyme exhibits a highly restricted substrate specificity and is able to cleave hydroxyquinol (Km for hydroxyquinol as a substrate was 1.2 μM, Vmax 55 U/mg, Kcat 57 s−1 and Kcat/Km 47.5 μM s−1), 6-chloro- and 5-chlorohydroxyquinol. Different substituted catechols and hydroquinones are not substrates for this enzyme. This enzyme appears to be a dimer with two identical 37-kDa subunits. Protein and iron analyses indicate an iron stoichiometry of 1 iron/65 kDa homodimer, α2 Fe. Both the electronic absorption spectrum which shows a broad absorption band with a maximum at 450 nm and the electron paramagnetic resonance spectra are consistent with a high-spin iron(III) ion in a rhombic environment typical of the active site of intradiol cleaving enzymes. 相似文献
15.
Robert J. Deschenes Dana C. Hilt Judith K. Marquis Henry G. Mautner 《生物化学与生物物理学报:生物膜》1981,641(1):166-172
Tb3+, a fluorescent trivalent cation with physicochemical properties similar to Ca2+, binds to peripheral nerve membrane vesicles prepared from the walking leg nerve bundle of the lobster (Homarus americanus). Saturable binding is measured for at least two classes of binding site. Bound Tb3+ can be displaced by other cations in the order: Ca2+ > Mg2+ = Zn2+ > NH4+. The binding of Tb3+ to the lower affinity site (KD(app) = 6.0 μM) is inhibitable by Na+, Mg2+ and Ca2+, whereas the higher affinity site (KD(app) = 2.2 μM) is only sensitive to Ca2+. Using this spectral probe the role of Ca2+ in peripheral nerve membrane function can be investigated. 相似文献
16.
We have examined hepatic microsomes prepared from phenobarbital-treated animals for the presence of the inducer and found that significant concentrations of the drug remain bound even after a number of washings. The bound drug is at least 70% unmetabolized and interferes with the in vitro binding of ethylmorphine and phenobarbital to the type I binding site but is not bound to the type I site since the Ks is 0.2–6.4 μm whereas the Ks for type I binding of phenobarbital is 103 μm. The high affinity site was not observed in microsomes from control animals either spectrophotometrically or radiometrically. The bound drug can be removed by bovine serum albumin to give a reverse pseudo-type I spectrum with a peak at 412 nm rather than the 425 nm observed for the trough of the typical type I spectrum. These data suggest that induction with phenobarbital may alter the spectral properties of cytochrome P-450 and hence care should be taken in comparing spectral data between microsomes from phenobarbital-treated and control animals. 相似文献
17.
P. V. Ershov Yu. V. Mezentsev E. O. Yablokov L. A. Kaluzhskiy A. V. Florinskaya A. V. Svirid A. A. Gilep S. A. Usanov A. E. Medvedev A. S. Ivanov 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2018,12(2):130-135
Cytochrome P450-dependent monooxygenase systems exist basically in all living organisms, where they perform various important functions. The coordinated functioning of these systems involves many proteins participating in different protein-protein interactions (PPI). Previously, we have found that the endogenous non-peptide bioregulator isatin (indoledione-2,3), synthesized from indole by means of certain cytochromes P450 (e.g. P450 2E1, P450 2C19, P450 2A6) regulates affinity of some PPI. In this study, an attempt has been undertaken to register a direct interaction of isatin with a set of different proteins related to the functioning of cytochrome P450-dependent monooxygenase systems: five isoforms of cytochromes P450, two isoforms of cytochrome b5, cytochrome P450 reductase, adrenodoxin, adrenodoxin reductase and ferrochelatase. The study has shown high affinity specific binding of isatin only to cytochromes P450 (the equilibrium dissociation constant (Kd) is about 10–8 M). 相似文献
18.
Zufan Araya Kjell Wikvall 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》1999,1438(1):47-54
The aim of the present study was to identify the enzymes in human liver catalyzing hydroxylations of bile acids. Fourteen recombinant expressed cytochrome P450 (CYP) enzymes, human liver microsomes from different donors, and selective cytochrome P450 inhibitors were used to study the hydroxylation of taurochenodeoxycholic acid and lithocholic acid. Recombinant expressed CYP3A4 was the only enzyme that was active towards these bile acids and the enzyme catalyzed an efficient 6α-hydroxylation of both taurochenodeoxycholic acid and lithocholic acid. The Vmax for 6α-hydroxylation of taurochenodeoxycholic acid by CYP3A4 was 18.2 nmol/nmol P450/min and the apparent Km was 90 μM. Cytochrome b5 was required for maximal activity. Human liver microsomes from 10 different donors, in which different P450 marker activities had been determined, were separately incubated with taurochenodeoxycholic acid and lithocholic acid. A strong correlation was found between 6α-hydroxylation of taurochenodeoxycholic acid, CYP3A levels (r2=0.97) and testosterone 6β-hydroxylation (r2=0.9). There was also a strong correlation between 6α-hydroxylation of lithocholic acid, CYP3A levels and testosterone 6β-hydroxylation (r2=0.7). Troleandomycin, a selective inhibitor of CYP3A enzymes, inhibited 6α-hydroxylation of taurochenodeoxycholic acid almost completely at a 10 μM concentration. Other inhibitors, such as α-naphthoflavone, sulfaphenazole and tranylcypromine had very little or no effect on the activity. The apparent Km for 6α-hydroxylation of taurochenodeoxycholic by human liver microsomes was high (716 μM). This might give an explanation for the limited formation of 6α-hydroxylated bile acids in healthy humans. From the present results, it can be concluded that CYP3A4 is active in the 6α-hydroxylation of both taurochenodeoxycholic acid and lithocholic acid in human liver. 相似文献
19.
Michael J. Fasco Lawrence J. Piper Laurence S. Kaminsky 《Archives of biochemistry and biophysics》1977,182(2):379-389
The R and S enantiomers of the anticoagulant, warfarin, are metabolized to a series of monohydroxylated products by rat hepatic cytochrome P-450. The patterns of metabolites are a function of the warfarin enantiomer used and of the induction of the microsomal enzymes by phenobarbital (PB) and 3-methylcholanthrene (MC). We have studied the binding of R and S warfarin to cytochrome P-450 by difference spectrometry to probe the heterogeneity of cytochrome P-450 and to determine the role of this heterogeneity in the production of the patterns of warfarin metabolites. Uninduced cytochrome P-450 yielded modified type II spectra with R and S warfarin with equivalent binding constants, Ks = 1.50 mM. PB-induced cytochrome P-450 yielded modified type II spectra which varied biphasically with warfarin concentration with Ks(S) = 0.24 and 0.07 mm; Ks(R) = 0.79 and 0.12 mM. MC induction and 2-allyl-2-isopropylacetamide treatment yielded microsomes markedly enriched for cytochrome P-448 which, with R warfarin, yielded a type I spectrum, Ks = 0.24 mM, and with S warfarin a modified type II spectrum with Ks = 0.11 mM. The effects of the type I compound, hexobarbital, the type II compound, imidazole, or the opposite enantiomer to that being studied on the binding spectra of R and S warfarin to the variously induced cytochromes P-450 were investigated as an aid to elucidating the mode of interaction of cytochrome P-450 with warfarin. In all cases, prior binding of R or S warfarin influenced the binding of the opposite enantiomer. We conclude from these results that R and S warfarin bind to two separate forms of PB-induced cytochrome P-450 and two separate forms of MC-induced cytochrome P-448, all of which differ from uninduced cytochrome P-450. The variety of monohydroxylated metabolites of R and S warfarin is probably a consequence of the interactions with these different forms of cytochrome P-450. 相似文献
20.
The relative reactivity of 3 naphthoquinones, which are feeding inhibitors for Periplaneta americana and Scolytus multistriatus, with each of 7 amino acids was measured by ultraviolet difference spectroscopy. Juglone (5-hydroxy-1,4-naphthoquinone), menadione(2-methyl-1,4-naphthoquinone) or 1,4-naphthoquinone produced difference spectra immediately upon mixing with cysteine, but not with valine, serine, glutamic acid, arginine, tryptophan or proline in phosphate buffer (pH 7.0). The Ks values for the reactions indicated that the affinities of 1,4-naphthoquinone (Ks = 4.4 · 10−4M) and juglone (Ks = 8.3 · 10−4M) for cysteine were comparable, but were both approx. 10 times greater than that for menadione (Ks = 3.2 · 10−3M). The extinction coefficient of the complex formed by cysteine with juglone (A = 3.448 · 10−1M) was approx. 2 times greater than that of 1,4-naphthoquinone (A = 1.290 · 10−1M) or menadione (A = 1.176 · 10−1 M). The importance of these results to explaining the mechanism of chemoreception in P. americana and S. multistriatus is discussed. 相似文献