重金属Hg^2＋离子作用于PSⅡ 23 kD外周蛋白的光谱学研究

23kD蛋白是光系统Ⅱ（PSⅡ）的外周蛋白,具有增加Ca^2＋和Cl^-的结合以维持放氧活性的作用。本文借助内源荧光光谱和紫外吸收差谱考察了23kD蛋白与Hg^2＋的相互作用。所得结果表明：低浓度的Hg^2＋离子（〈10μM）引起23kD蛋白的荧光淬灭,淬灭程度与Hg^2＋离子浓度相关;进一步分析显示,Hg^2＋离子对23kD蛋白色氨酸荧光的淬灭属于静态淬灭。紫外吸收差谱可以检测到明显的配体向金属进行电荷转移的谱带（LMCT band）。随着Hg^2＋浓度的增加,LMCT带的强度逐渐增强。分析显示23kD蛋白只存在一类Hg^2＋离子结合位点。     

重金属Hg2+离子作用于PSII 23 kD外周蛋白的光谱学研究

23 kD蛋白是光系统II(PSII)的外周蛋白,具有增加Ca2 和C l-的结合以维持放氧活性的作用。本文借助内源荧光光谱和紫外吸收差谱考察了23 kD蛋白与Hg2 的相互作用。所得结果表明:低浓度的Hg2 离子(<10μM)引起23 kD蛋白的荧光淬灭,淬灭程度与Hg2 离子浓度相关;进一步分析显示,Hg2 离子对23 kD蛋白色氨酸荧光的淬灭属于静态淬灭。紫外吸收差谱可以检测到明显的配体向金属进行电荷转移的谱带(LMCT band)。随着Hg2 浓度的增加,LMCT带的强度逐渐增强。分析显示23 kD蛋白只存在一类Hg2 离子结合位点。     

根霉超氧化物歧化酶同工酶类型的鉴别

利用ＫＣＮ、Ｈ２Ｏ２睡ＳＤＳ的选择性反应引起的ＳＯＤ同工酶谱带变化即可鉴别精 抽提液中的ＳＯＤ同工酶类型,用这种方法对五株不同种根霉的鉴别实验表明,它们均不同程度地含有Ｃｕ,Ｚｎ型和Ｍｎ型两各ＳＯＤ,前者约点８０％左右,后者只占２０％左右,用邻苯三酚自氧化法对样品中ＳＯＤ活性测定结果与在同工酶谱上的鉴别结果基本一致。     

根霉超氧化物歧化酶同工酶类型的鉴别

利用ＫＣＮ、Ｈ_２Ｏ２和ＳＤＳ的选择性反应引起的ＳＯＤ同工酶谱带变化即可鉴别粗抽提液中的ＳＯＤ同工酶类型;用这种方法对五株不同种根霉的鉴别实验表明,它们均不同程度地含有Ｃｕ,Ｚｎ型和Ｍｎ型两种ＳＯＤ,前者约占８０％左右,后者只占２０％左右。用邻苯三酚自氧化法对样品中ＳＯＤ活性测定结果与在同工酶谱上的鉴别结果基本一致。     

Zn^2^＋对细菌铁蛋白形成的影响

用酶联免疫法（ＥＬＩＳＡ）检测了Ｚｎ＾２＾＋对细菌铁蛋白合成的影响。Ｚｎ＾２＾＋抑制Ｂ．Ｆ抑制Ｂ．Ｆ合成,而在高Ｚｎ＾２＾＋培养液中生长的菌体其Ｂ．Ｆ分子中Ｚｎ＾２＾＋含量比低Ｚｎ＾２＾＋条件下的高二部。Ｂ．Ｆ分子中的Ｚｎ＾２＾＋含量比低Ｚｎ＾２＾＋条件下的高二倍。Ｂ．Ｆ分子中的Ｚｎ＾２＾＋能转运出来激活碱性磷酸酶的活性。     

D1／D2／Cyt—559复合物的共振拉曼光谱

光系统Ⅱ（ＰＳⅡ）反应中心Ｄ１／Ｄ２／Ｃｙｔｂ－５５９复合物在４８８ｎｍ处激发的共振拉曼谱显示４个主要谱带,其峰位分别在１５３２（ｖｌ）１１６５（ｖ２）１０１０（ｖ３）和９７０（ｖ４）ｃｍ＾－１处,表明ＰＳⅡ反应中心结合的β－胡萝卜素分子是全反式构型。Ｄ１／Ｄ２／Ｃｙｔｂ－５５９复合物在色素抽提液的拉曼光谱也显示４个主要的拉曼峰,其中ｖ４谱带的强度急剧下降,说明ＰＳⅡ反应中心内部结合的β－胡萝卜素     

铜锌超氧化物歧化酶与过氧化氢反应中羟自由基的形成

应用脱氧核糖降解法研究了离体条件一Ｃｕ,Ｚｎ－ＳＯＤ与Ｈ２Ｏ２反应产生．ＯＨ,并对其机理进行了探讨。Ｈ２Ｏ２可使Ｃｕ,Ｚｎ－ＳＯＤ失活,在失活过程中有．ＯＨ产生。甲酸钠和苯甲酸钠均能不同程度地保护Ｃｕ,Ｚｎ－ＳＯＤ和降低Ｈ２Ｏ２与ＣｕＺｎ－ＳＯＤ反应中。ＯＨ的产额,热失活ＳＯＤ也可和Ｈ２Ｏ２反应生成ＯＨ,且效能高于活性Ｃｕ,Ｚｎ－ＳＯＤ;     

马尾松毛虫核型多角体病毒包涵体的表面增强拉曼光谱研究

获得了马尾松毛虫核型多角体病毒（ＤｐＮＰＶ）包涵体在Ａｇ胶中的ＳＥＲＳ谱。包涵体是通过膜蛋白的ＣＯＯ－和ＮＨ２基团吸附在Ａｇ表面上,Ｔｒｐ和Ｐｈｅ残基侧链接近Ａｇ表面,Ｔｙｒ残基侧链远离Ａｇ表面,ＳＥＲＳ谱中没出现ＡｍｉｄｅⅠ和ＡｍｉｄｅⅢ带。强的低波数谱带（２３８ｃｍ－１）的出现表明包涵体与Ａｇ表面发生了明显的化学吸附。     

外源^65Zn进入土壤后的扩散及存在形态

利用６５Ｚｎ示踪和化学连续分级技术研究了外源Ｚｎ在褐土中的存在形态。结果表明交换态Ｚｎ（ＥＸ－Ｚｎ）和碳酸盐结合态Ｚｎ（ＣＡＲＢ－Ｚｎ）含量约占土壤总Ｚｎ量的６７－７２％．Ｚｎ投加量增加,土壤Ｚｎ的强度因数增大．反之,Ｚｎ的矿物形态增高．土壤水分４％和１００％时,土壤中Ｚｎ的扩散系数Ｄ分别是７．９×１０－８和６．６×１０－６ｃｍ２·ｓ－１．土壤水分含量在３０％和７０％时,Ｚｎ在施入点的平均停留时间分别是９ｄ和４ｄ．Ｚｎ的扩散程度（σ２）也随之从０增大到０．３７１．     

癌症病人血液的荧光光谱研究

研究了癌症病人全血、血浆和血红蛋白的激发波长—发射波长—荧光光强三维光谱,观察到特征谱带625nm的荧光本原物质存在于血红蛋白部分,比较了十五种癌症(28例),十例非癌症病人和健康人血液在625nm带的荧光强度.     

Excitation energy transfer study of the spatial relationship between the carbonyl and metal cofactors in pig plasma amine oxidase

9-Hydrazinoacridine irreversibly labeled pig plasma amine oxidase by covalent attachment to the active carbonyl cofactor. The visible absorption spectrum of the modified protein displays new absorption bands at 495 and 525 nm. Its emission spectrum exhibited maxima at 415 and 440 nm. In addition, both absorption and emission spectra were insensitive to pH changes between 6 and 10. Phase modulation fluorometry was used to determine fluorescence lifetimes of Zn2+- and Co2+-substituted acridinyl plasma amine oxidase. Energy transfer efficiency was 22%; the distance separating the Co2+ ion (in the copper binding site) and the acridine moiety (the amine substrate binding site) ranges between 11.7 and 14.7 A. This work defines the proximity of the metal and substrate (and hence the carbonyl cofactor) and precludes any direct interaction between Cu2+ and pyrroloquinoline quinone or between Cu2+ and the substrate.     

Some dicopper complexes of benzimidazole-containing ligands.

Dicopper complexes of the following benzimidazole-containing ligands have been studied as possible models for the active site of hemocyanin: EDTB (N,N,N',N'-tetrakis-(2-benzimidazolylmethyl)-1,2-ethanediamine), EGTB (1,1,10,10-tetrakis-(2-benzimidazolylmethyl)-1,10-diaza-4,7- dioxadecane), and MEGTB (1,1,10,10-tetrakis-(1-methylbenzimidazol-2-y lmethyl)-1,10-diaza-4,7-dioxadecane). The initial oxygenation product of Cu2(EDTB)(ClO4)2 in Me2SO gives optical absorption maxima at 315 nm (epsilon = 3750 M-1 cm-1) and 690 nm (epsilon = 100 M-1 cm-1). The fluorescence emission intensities of Cu2(EDTB)(ClO4)2 at 400 and 700 nm (excitation at 350 nm) decreases rapidly on exposure to air. This suggests oxidation of Cu2(I) to Cu2(II). The x-ray absorption edge spectra suggest that both coppers in the oxygenation product, analyzed as Cu2(EDTB)(ClO4)2(O).3H2O, are Cu(II). From spectrophotometric titration of Cu2(MEGTB)Cl4 with azide, formation constant of the Cu2(MEGTB)N3Cl3 complex has been obtained. Data from cyclic voltammetry experiments suggest that in the presence of azide, Cu(II)(N3)Cu(II) species is present.     

Two new supermolecular structures of organic-inorganic hybrid compounds: [Zn(phen)(SO4)(H2O)2]n and [Cu(phen)(H2O)2] · SO4 (phen = 1,10-phenanthroline)

Two new organic-inorganic hybrid compounds [Zn(phen)(SO₄)(H₂O)₂]_n (1) and [Cu(phen)(H₂O)₂] · SO₄ (2) have been prepared by conventional aqueous solution synthesis and characterized by single-crystal X-ray diffraction, IR spectroscopy, thermal gravimetric analysis (TGA) and fluorescent spectroscopy. In compound 1, the sulfate group adopts bidentate mode to coordinate with two Zn(II) ions to form one-dimensional polymer. The one-dimensional polymers are further linked together via the intermolecular hydrogen-bonding and π-π stacking interactions to form a 3D supramolecular framework. Compound 2 is build up of discrete [Cu(phen)(H₂O]²⁺ cations and SO₄²⁻ anions to form a three-dimensional framework via hydrogen-bonding and π-π stacking interactions. Furthermore, the luminescent properties of both 1 and 2 were studied. The complexes 1 and 2 excited at 280 nm wavelength produced characteristic luminescence features, arising maybe due to the π-π transitions.     

Electron paramagnetic resonance study of the active site of copper-substituted human glyoxalase I

Zn2+ in native glyoxalase I from human erythrocytes can be replaced by Cu2+, giving an inactive enzyme. Cu2+ was demonstrated to compete with the activating metals Zn2+ and Mn2+, indicating a common binding site on the enzyme for these metal ions. The electron paramagnetic resonance (EPR) spectra of 63Cu(II) glyoxalase I at 77 K and of its complexes with glutathione and some glutathione derivatives are characteristic of Cu2+ in an elongated octahedral coordination (g parallel = 2.34, g perpendicular = 2.09, and A parallel = 14.2 mT). The low-field bands of the free enzyme are asymmetric and become symmetrical upon addition of glutathione or S-(p-bromobenzyl)glutathione but not S-(D-lactoyl)glutathione. The results indicate the existence of two conformations of Cu(II) glyoxalase I, in agreement with the effects caused by these compounds on the protein fluorescence. The copper hyperfine line at low field in the EPR spectrum of the S-(p-bromobenzyl)glutathione complex of 63Cu(II) glyoxalase I shows a triplet structure, indicative of coupling to one nitrogen ligand in the equatorial plane. Similar results were obtained with the glutathione complex. By addition of the spectrum of the S-(p-bromobenzyl)glutathione complex and a spectrum corresponding to two nitrogen ligands with two different coupling constants, a good fit was obtained for the low-field region of the asymmetric spectrum of free 63Cu(II) glyoxalase I. The first two spectra are assumed to correspond to two separate conformational states of the enzyme. The results demonstrate that at least one nitrogen ligand is involved in the binding of Cu2+.(ABSTRACT TRUNCATED AT 250 WORDS)     

Spectroscopic and kinetics studies of the inhibition of pig kidney diamine oxidase by anions

The role of copper in pig kidney diamine oxidase has been probed by examining the effects of potential Cu(II) ligands on the spectroscopic and catalytic properties of the enzyme. In the presence of azide and thiocyanate, new absorption bands are evident at 410 nm (epsilon = 6300 M-1 cm-1) and 365 nm (epsilon = 3000 M-1 cm-1), respectively. These bands are assigned as ligand-to-metal charge-transfer transitions, N3-/SCN- leads to Cu(II). One anion/Cu(II) is coordinated in an equitorial position. Anion binding can be completely reversed by dialysis. The equilibrium constants for diamine oxidase-anion complex formation are 134 M-1 (N3-) and 55 M-1 (SCN-). Azide and thiocyanate are linear uncompetitive inhibitors with respect to the amine substrate when O2 is present at saturating concentrations. Taken together, the data are consistent with a functional role for Cu(II) in diamine oxidase catalysis.     

Ternary Cu(II) complexes, Cu(H(-1)L)(ACys-) and Cu(H(-2)L)(ACys-); L = peptides, ACys- = N-acetyl-cysteinate. Analogous complexes to the intermediates in the transport of Cu(II) from Cu(H(-2)L) to cysteine

The Cu(II) in Cu(H(-2)L) has been postulated to be successively transported to cysteine (Cys) as follows; Cu(H(-2)L) <==> Cu(H(-2)L)(Cys*-) <==> Cu(H(-1)L)(Cys*-) --> Cu(H(-1)L)(Cys-), where Cys*- denotes the monodentate Cys-. N-acetyl-cysteinate (ACys-) complexes Cu(H(-2)L)(ACys-) and Cu(H(-1)L)(ACys-), having similar coordination modes to Cu(H(-2)L)(Cys*-) and Cu(H(-1)L)(Cys*-), respectively, exhibited the S --> Cu(II) charge transfer absorption at 325-355 nm and the d-d absorption at 530-610 nm. A linear interrelation existed between the energies of the CD and d-d absorptions. Cu(H(-2)L)(ACys-) were in rapid equilibrium with Cu(H(-1)L)(ACys-). Upon forming the ternary complex, pK(c2) of the parent Cu(H(-1)L) was raised to more than 1.0. The formation constants (K) of the Cu(H(-1)L)(ACys-) species from Cu(H(-1)L) were bigger than those of Cu(H(-2)L)(ACys-) from Cu(H(-2)L). The linear free-energy relationship existed between the free-energy change (deltaG) and the entropy change (deltaS) for the ternary complex formation. The rate constants (k1+) for the Cu(H(-1)L)(Cys-) formation closely correlated with the K values for Cu(H(-2)L)(ACys-). The ternary complexes containing ACys are considered to be analogous complexes to the intermediates in the transport of Cu(II) from peptides to cysteine.     

E.s.r., visible and SOD studies of imidazolate bridged Cu(2)(II,II), Cu(II)Zn(II) and Cu(II)Ni(II) complexes with pentamethyldiethylenetriamine as capping ligand: a plausible model for superoxide dismutase

X-band e.s.r. and electronic spectra of imidazolate bridged homobinuclear Cu-Cu complex, [(PMDT)Cu-Im-Cu(PMDT)](ClO(4))(3) and heterobinuclear Cu-Zn and Cu-Ni complexes, viz. [(PMDT)Cu-Im-Zn(PMDT)](ClO(4))(3), [(PMDT)Cu-Im-Ni(PMDT)] (ClO(4))(3), where PMDT=pentamethyldiethylenetriamine, Im=Imidazolate ion and related mononuclear complexes, [(PMDT)Cu(OH(2))](2+) and [(PMDT)Cu(ImH)](2+) have been described. Superoxide dismutase activities of these complexes have also been measured.     

Resonance Raman spectra of the copper-sulfur chromophores in Achromobacter cycloclastes nitrite reductase

Resonance Raman spectroscopy at ambient temperature and 77 K has been used to probe the structures of the copper sites in Achromobacter cycloclastes nitrite reductase. This enzyme contains three copper ions per protein molecule and has two principal electronic absorption bands with lambda max values of 458 and 585 nm. Comparisons between the resonance Raman spectra of nitrite reductase and blue copper proteins establish that both the 458 and 585 nm bands are associated with Cu(II)-S(Cys) chromophores. A histidine ligand probably is also present. Different sets of vibrational frequencies are observed with 457.9 nm (ambient) or 476.1 nm (77 K) excitation as compared with 590 nm (ambient) or 593 nm (77 K) excitation. Excitation profiles indicate that the 458 and 585 nm absorption bands are associated with separate [Cu(II)-S(Cys)N(His)] sites or with inequivalent and uncoupled cysteine ligands in the same site. The former possibility is considered to be more likely.     

Impaired glucose homeostasis and mitochondrial abnormalities in offspring of rats fed a fat-rich diet in pregnancy

This study examined the role of heating on oxidative stress and muscle mass in immobilized limbs. Rats were divided into three groups (n = 9/group): a control group (Con), an immobilized group (Im), and an immobilized and heated group (ImH). Rats were immobilized in the plantarflexed position for 8 days. The core temperature of the ImH group was elevated to 41-41.5 degrees C on alternating days and maintained for 30 min before cooling. On day 8, both heat shock protein 25 (HSP25) and HSP72 were markedly elevated in the ImH compared with the Im group, whereas results in the Im group were not different from Con. Most notably, the ImH group had significantly larger solei compared with the Im group, which were less than those shown in the Con group. Furthermore, immobilization alone caused a significant increase in oxidative damage, and the addition of heating to immobilization significantly reduced oxidative damage. In an effort to further identify the cause of this protective effect, antioxidant enzyme activities were assessed. CuZnSOD was sharply elevated in Im compared (P < 0.025) with that in the Con and reduced in the ImH group compared with that in the Im group (P < 0.025). Catalase was elevated 8% (P < 0.025) in the Im group compared with the Con group and was similar to the ImH group. Glutathione peroxidase, glutathione reductase, and MnSOD did not differ between groups. These data indicate that heating provides protection against oxidative stress and preserves muscle mass during disuse atrophy. These data also suggest that antioxidant protection is not conferred via antioxidant enzymes, and HSPs may play an important role.     

L-SOD发挥活性作用的可能机制

氧化剂、还原剂处理前后,Ｌ－ＳＯＤ的活性及紫外光谱发生变化,Ｈ２Ｏ２使Ｆｅ（Ⅲ）吸收增强,同时钝化Ｌ－ＳＯＤ的活性;加入保险粉后,Ｌ－ＳＯＤ重新活化,Ｆｅ（Ⅲ）吸收减弱．ＮＥＭ封闭Ｃｙｓ后,Ｌ－ＳＯＤ紫外吸收谱发生变化,且活性减弱．说明Ｆｅ辅基及Ｃｙｓ是活性发挥的必需基团．