硒代半胱氨酸(selenocysteine):密码表中第二十一个氨基酸?

已知所有的氨基酸中,只有遗传密码表内的20种基本氨基酸才能在核糖体中直接掺入肽链;但最近发现于原核、真核生物中的含硒酶里的硒代半胱氨酸(Se-Cys)似亦有此特点。生化、遗传实验均表明Se-Cys对应于终止密码子UGA;相应的tRNA(95bp)基因已经找到。但其转录产物上所携的Se-Cys很可能由原先携带着的Ser经O-磷酰Ser而来。上述发现显示了UGA作为有义密码子的保守性:也许它正处于从有义密码子变为无义密码子的进化过程中。     

硒代磷酸合成酶的研究进展

硒代磷酸合成酶（ＳＰＳ）的催化产物硒代磷酸（ＳＰ）是合成硒蛋白所必需的活性硒供体,所以ＳＰＳ可能对硒蛋白合成的调控起重要作用。近来发现ＳＰＳ广泛分布于哺乳动物、细菌和古细菌。ＳＰＳ菌化如下反应：ＡＴＰ＋硒化合物＋Ｈ２Ｏ→ＳＰ＋ＡＭＰ＋正磷酸。ＡＴＰ是该酶的特异性底物,ＡＭＰ是其竞争性抑制剂。该酶表现催化活性必需Ｍｇ＾２＋和一阶阳离子Ｋ＾＋等。     

氨基酸叶面硒肥对铁皮石斛富硒及多糖含量的影响

研究不同浓度氨基酸叶面硒肥(CK、1200倍液、900倍液和600倍液)对智能温室人工栽培红杆铁皮石斛(Dendrobium officinale)枝条(即鲜条)富硒、抽芽和多糖含量的影响。结果表明,氨基酸硒肥各浓度处理均极显著提高鲜条硒和多糖含量。鲜条硒含量与氨基酸硒肥浓度呈正相关,且各处理间达极显著差异。鲜条多糖含量在氨基酸硒肥浓度较低(CK、1200、900倍液)时呈正相关,并在氨基酸硒肥900倍液时达最高(35.213 g·kg-1),之后呈递减趋势。氨基酸硒肥稀释达900倍时,90%以上鲜条已抽出3片韧叶,表明氨基酸硒肥明显促进鲜条生长。因此,环保型氨基酸硒肥900倍液是铁皮石斛营养积累前期较适宜浓度。     

用微生物酶合成旋光的硫和硒氨基酸

<正> 自然界中存在各种硫化合物,而且大多数是生理活性的。甲硫氨酸、半胱氨酸和其它一些硫氨基酸尤其起着重要的代谢作用。例如,L-甲硫氨酸不仅是一种必需氨基酸,而且是通过中间体S-腺苷-L-甲硫氨酸的各种转甲基酶系中的主要供体。它还是多胺和一种植物激素—乙烯的重要前体。大量的DL-甲硫氨酸广泛用作黄豆粉和其它缺乏硫氨基酸的饲料的添加剂。甲硫氨酸、半胱氨酸和其它氨基酸还是药物、化妆品和化学品的很重要的原料。目前利用细菌可以由简单的碳、氨源生产各种L-氨基酸,包括谷氨酸和赖氨酸。但用微生物方法生产硫氨基酸则很少获得成功。     

含硫和含硒氨基酸及其光学异构体的气相色谱分离和分析

<正> 这篇综述的写作构思有三:一是时至今天,含硫特别是含硒氨基酸的准确分析仍然面临不少难题;二是硒蛋白及组成它的含硒氨基酸是当前生物化学的热门课题之一,例如有人提出硒半胱氨酸可能是构成生命遗传密码的第21种氨基酸;三是近10多年来国内外还未见有这方面的专门综述。     

谷胱甘肽过氧化物酶的硒代半胱氨酸插入元件

真核生物将硒代半胱氨酸插入蛋白质必需硒代半胱氨酸插入元件（ＳＥＣＩＳ）的参与,后者位于硒蛋白ｍＲＮＡ的３′非翻译区．采用ＲＮＡ折叠程序对１５个谷胱甘肽过氧化物酶基因进行计算机处理发现,其可能的ＳＥＣＩＳ中都具有３段保守碱基ＡＵＧＡ－Ａ（Ｇ）ＡＡ－ＧＡ．根据Ａ（Ｇ）ＡＡ位于顶环或者顶环上游５′臂的突环上,可将ＳＥＣＩＳ分为Ⅰ型和Ⅱ型结构     

Selenium containing amino acids and proteins in marine algae

The unicellular marine algae, Dunaliella primolecta Butcher, Chlorella sp. and Porphyridium cruentum (S.F. Grey) were grown in artificial sea water containing a sublethal concentration of selenite, 10^?2 g Se/1. Both free-and protein-bound seleno-amino acids were identified. The initial steps of selenium incorporation seem to involve the use of the sulfur enzymatic machinery resulting in the replacement of some of the sulfur by selenium in both free amino acids and proteins. At relatively low selenium concentrations, selenium-specific enzymes seem to be in operation.     

Analysis of free amino acids in mammalian brain extracts

An optimized method for analysis of free amino acids using a modified lithium-citrate buffer system with a Hitachi L-8800 amino acid analyzer is described. It demonstrates clear advantages over the sodium-citrate buffer system commonly used for the analysis of protein hydrolysates. A sample pretreatment technique for amino acid analysis of brain extracts is also discussed. The focus has been placed on the possibility of quantitative determination of the reduced form of glutathione (GSH) with simultaneous analysis of all other amino acids in brain extracts. The method was validated and calibration coefficient (K _GSH) was determined. Examples of chromatographic separation of free amino acids in extracts derived from different parts of the brain are presented.     

Analysis of amino acids and carbohydrates in green coffee

The analysis of carbohydrates and amino acids in green coffee is of the utmost importance since these two classes of compounds act as precursors of the Maillard reaction during which the colour and aroma are formed. During the course of the Maillard reaction potentially harmful substances like acrylamide or 5-hydroxymethyl-furfural accrue as well. The carbohydrates were analysed by anion-exchange chromatography with pulsed amperometric detection and the amino acids by reversed phase chromatography after derivatization with 6-amino-quinolyl-N-hydroxysuccinimidyl carbamate and fluorescence detection. Both methods had to be optimized to obtain a sufficient resolution of the analytes for identification and quantification. Sucrose is the dominant carbohydrate in green coffee with a concentration of up to 90 mg/g (mean = 73 mg/g) in arabica beans and significantly lower amounts in robusta beans (mean=45 mg/g). Alanine is the amino acid with the highest concentration (mean = 1200 microg/g) followed by asparagine (mean = 680 microg/g) in robusta and 800 microg/g and 360 microg/g in arabica respectively. In general, the concentration of amino acids is higher in robusta than in arabica.     

Analysis of amino acids in individual wheat embryonic protoplast

Summary. Amino acids analysis in single wheat embryonic protoplast was performed using capillary electrophoresis equipped with laser-induced fluorescence (CE-LIF), combination with tissue culture technique. Reagent fluorescein isothiocyanate (FITC) was introduced into living protoplasts by electroporation for intracellular derivatization. A special osmotic buffer (0.6 mol/L mannitol, 5 mmol/L CaCl₂) was used to keep the osmotic balance of embryonic protoplasts during the protoplasts derivatization. After completion of the derivatization reaction in the protoplasts, a single protoplast was drawn into the capillary tip by electroosmotic flow. Then a 0.1 M NaOH lysing solution was injected by diffusion. The derivatized amino acids were separated by capillary electrophoresis and detected by laser-induced fluorescence detection after the protoplast was lysed Nine amino acids were quantitatively and qualitatively determined and compared in lysate and single protoplast of wheat embryonic cells respectively, with mean concentrations of amino acids ranging from 2.68×10⁻⁵ mol/L to 18.18×10⁻⁵ mol/L in single protoplast.     

Transformation of some hydroxy amino acids to other amino acids.

It has been observed that beta-hydroxy-alpha-amino acids are transformed into other amino acids, when heated in dilute solutions with phosphorous acid, phosphoric acid or their ammonium salts. It has been shown that as in the case of previously reported glycine-aldehyde reactions, glycine also reacts with acetone to give beta-hydroxyvaline under prebiologically feasible conditions. It is suggested, therefore, that the formation of beta-hydroxy-alpha-amino acids and their transformation to other amino acids may have been a pathway for the synthesis of amino acids under primitive earth conditions.     

Selenium derivatization of nucleic acids for crystallography

The high-resolution structure of the DNA (5′-GTGTACA-C-3′) with the selenium derivatization at the 2′-position of T2 was determined via MAD and SAD phasing. The selenium-derivatized structure (1.28 Å resolution) with the 2′-Se modification in the minor groove is isomorphorous to the native structure (2.0 Å). To directly compare with the conventional bromine derivatization, we incorporated bromine into the 5-postion of T4, determined the bromine-derivatized DNA structure at 1.5 Å resolution, and found that the local backbone torsion angles and solvent hydration patterns were altered in the structure with the Br incorporation in the major groove. Furthermore, while the native and Br-derivatized DNAs needed over a week to form reasonable-size crystals, we observed that the Se-derivatized DNAs grew crystals overnight with high-diffraction quality, suggesting that the Se derivatization facilitated the crystal formation. In addition, the Se-derivatized DNA sequences crystallized under a broader range of buffer conditions, and generally had a faster crystal growth rate. Our experimental results indicate that the selenium derivatization of DNAs may facilitate the determination of nucleic acid X-ray crystal structures in phasing and high-quality crystal growth. In addition, our results suggest that the Se derivatization can be an alternative to the conventional Br derivatization.     

Solubilities of amino acids

The Mettler/Paar precision density meter DMA-02D has been used to determine the concentration of saturated solutions of amino acids at 20.0, 25.0, and 29.8 °C. The technique has proven itself an elegant and precise method. The solubilities of all of the amino acids with the exceptions of proline, lysine, and cystine have been measured. The Gibbs free energies of transfer from saturated water solution to 1M Na₂SO₄ and to 1M Gu·HCL along with the van't Hoff heats and entropies have been calculated. The van't Hoff heats have been compared with the calorimetrically determined heats for some of the amino acids. The Lumry-Rajender relation between the entropy and heats has been observed. The process of transfer of the amino acids from water to the solvents is primarily enthalpic rather than entropic.