Structural and functional characterization of peptide-beta2m fused HLA-A2/MART1(27-35) complexes

The uses of soluble HLA class I/peptide complexes to monitor antigen reactive T cells are often hampered by their low-yield and high-cost production. As an alternative strategy, the peptide-beta(2)m fused, 2-component (2C) HLA class I/peptide complex has been developed, but its application is limited due to the lack of the comparison of its structural and functional characteristics with those of its conventional 3-component (3C) counterpart. In this study, we have demonstrated that the 2C and 3C HLA-A2/MART1(27-35) complexes have a similar chromatographical profile and comparable stability, but the former has 2.5 times higher yield and significantly higher binding ability with HLA-A2/MART1(27-35) complex-specific receptors than the latter. Furthermore, the 2C complex has a comparable ability to stimulate specific CTL proliferation, but appears to be more effective in eliciting the cytotoxicity of antigen-specific CTL, as compared to its 3C counterpart.     

Deciphering interactions of the aminoglycoside phosphotransferase(3′)‐IIIa with its ligands

Aminoglycoside phosphotransferase(3′)‐IIIa (APH) is the enzyme with broadest substrate range among the phosphotransferases that cause resistance to aminoglycoside antibiotics. In this study, the thermodynamic characterization of interactions of APH with its ligands are done by determining dissociation constants of enzyme–substrate complexes using electron paramagnetic resonance and fluorescence spectroscopy. Metal binding studies showed that three divalent cations bind to the apo‐enzyme with low affinity. In the presence of AMPPCP, binding of the divalent cations occurs with 7‐to‐37‐fold higher affinity to three additional sites dependent on the presence and absence of different aminoglycosides. Surprisingly, when both ligands, AMPPCP and aminoglycoside, are present, the number of high affinity metal binding sites is reduced to two with a 2‐fold increase in binding affinity. The presence of divalent cations, with or without aminoglycoside present, shows only a small effect (<3‐fold) on binding affinity of the nucleotide to the enzyme. The presence of metal–nucleotide, but not nucleotide alone, increases the binding affinity of aminoglycosides to APH. Replacement of magnesium (II) with manganese (II) lowered the catalytic rates significantly while affecting the substrate selectivity of the enzyme such that the aminoglycosides with 2′‐NH₂ become better substrates (higher V_max) than those with 2′‐OH. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 801–809, 2009. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com     

Potential molecular mechanisms for combined toxicity of arsenic and alcohol

Arsenic is a ubiquitous environmental factor that has been identified as a risk factor for a wide range of human diseases. Alcohol is clearly a toxic substance when consumed in excess. Alcohol abuse results in a variety of pathological effects, including damages to liver, heart, and brain, as well as other organs, and is associated with an increased risk of certain types of cancers. In history, arsenic-contaminated beers caused severe diseases. There are populations who are exposed to relatively high levels of arsenic in their drinking water and consume alcohol at the same time. In this focused review, we aim to discuss important molecular mechanisms responsible for arsenic toxicity and potential combined toxic effects of alcohol and arsenic.     

Conserved Chromosomal Location and Genomic Structure of Human and Mouse Fatty-Acid Amide Hydrolase Genes and Evaluation ofclasperas a Candidate Neurological Mutation

Fatty-acid amide hydrolase (FAAH) is a membrane-bound enzyme that degrades neuromodulatory fatty acid amides, such as oleamide and anandamide, and is expressed in the mammalian central nervous system. To evaluateFAAHgenes as candidates for neurogenetic diseases in humans and mice, we have mapped the loci in both species and have determined their intron–exon structures. The humanFAAHgene was mapped to region 1p34–p35, closely linked to D1S197 and D1S443, by using PCR analysis of somatic cell hybrid (SCH) and radiation hybrid mapping panels. Analysis of an SCH mapping panel and a mouse interspecific backcross panel has localized theFaahgene to the conserved syntenic region on mouse chromosome 4, close to the neurological mutationclasper. Faahgene rearrangements were excluded by Southern blot analysis of clasper DNA. No sequence abnormality was detected in PCR products containing the 15 exons and splice junctions of the mouseFaahgene. FAAH protein levels were normal in clasper mouse tissues as determined by enzyme activity assays and Western blotting.     

接种促生菌对尾巨桉—降香黄檀混作幼苗光合生理特性和生长的影响

该试验以盆栽尾巨桉和降香黄檀幼苗为材料,设置BM处理(尾巨桉接种巨大芽孢杆菌,降香黄檀不接种)、RJ处理(降香黄檀接种大豆根瘤菌,尾巨桉不接种)以及对照组(CK,尾巨桉和降香黄檀均不接菌),探究接种2种促生细菌对尾巨桉-降香黄檀混作幼苗的光合生理、生长和生物量积累及分配的影响,明确在混交体系中接种促生菌对促进植物生长的优势。结果显示:(1)BM处理显著降低尾巨桉的叶绿素含量(P<0.05),BM和RJ处理均提高了尾巨桉和降香黄檀的苗高、地径以及叶片的氮素含量、净光合速率、气孔导度、蒸腾速率,但降低了胞间CO_(2)浓度。(2)RJ处理显著提高了尾巨桉及降香黄檀叶片和全株生物量,BM处理仅显著提高降香黄檀根、茎、叶、全株的生物量和尾巨桉叶的生物量(P<0.05)。(3)各接菌处理条件下2种植物幼苗叶片净光合速率与其生物量呈显著正相关。研究表明,接种大豆根瘤菌和巨大芽孢杆菌均促进尾巨桉-降香黄檀混作幼苗的生理代谢,2种促生菌能通过增强光合作用来促进植株生物量的累积;从植株生物量变化来看,接种菌株的利他作用更明显。     

英国 Hampsfell 蕨菜草地生态系统的第一性生产量的研究

 4月中、下旬出现在英国的Hampsfell(英格兰的西北部)地面的蕨菜,以前两个月的生长速度最快,9月叶片达最大高度,但其重量高峰则出现在8月。地上部分生物量也以8月为最高,平均792克·米-2,其后逐渐下降,于11月地面部分枯死。地下部分的生物量季节变化不很明显,但随着地上部分的生长,地下部分的生物量也有所增长。土层深度是影响蕨菜生物量的最重要因子。蕨菜草地生态系统的第一性生产量平均为778克·米-2·年-1,在土深地段平均为918克·米-2·年-l,在浅土地段为525克·米-2·年-l。     

鲤、鲫肌肉水解氨基酸和游离氨基酸的初步研究

本文是对天津地区的两种重要的经济淡水鱼鲤、鲫肌肉水解氨基酸和游离氨基测定,并初步分析比较的结果。       

大鼠肝脏半乳糖凝聚素-3 cDNA分子多样性分析

半乳糖凝聚素-3(Galectin-3,Gal-3)是一种多功能蛋白,参与机体内的多种生命活动过程。为分析大鼠Rattus norvegicus gal-3 c DNA分子多样性,在分析Gen Bank注册的大鼠gal-3 c DNA序列的同时,从三只不同大鼠个体的肝脏第一链c DNA,共计9个独立实验中克隆gal-3开放阅读框OFR(Open reading frame)。分析结果表明,在已注册的序列中存在1个单核苷酸多态性(Single nucleotide polymorphism,SNP)。本实验从1-3号大鼠肝脏分别获得118、95和120个克隆,新发现1处缺失和5处SNP,共克隆17个不同的gal-3 ORF,其中仅1个克隆与已报告大鼠gal-3 c DNA的ORF完全相同。从1号和3号个体肝脏中分别克隆11和14个不同的gal-3 ORF,揭示大鼠gal-3 c DNA存在个体水平的分子多样性。