Specificities of Red Cell Membrane Sites Transporting Three Long Chain Fatty Acids

We studied the specificities of human red cell membrane bindings of three long chain fatty acids, palmitic- arachidonic- and oleic acid, using resealed membranes, ghosts. Previously estimated binding capacities, affinities and inside/outside distributions [6, 10, 11, 12], suggest separated binding sites. This possibility is explored by estimating the binding properties of one fatty acid in the presence of one or two of the others. Binding capacities, nmol g⁻¹ ghosts, of palmitic and arachidonic acid estimated simultaneously vs. separately are 27.4 ± 2.7 vs. 29.0 ± 2.1 (P < 0.6) and 6.5 ± 0.6 vs. 5.5 ± 0.5 (P < 0.2) respectively. The corresponding estimates for oleic- and palmitic acid are 36.5 ± 2.0 vs. 34.0 ± 2.2 (P < 0.4) and 28.4 ± 1.8 versus 29.1 ± 2.1 (P < 0.8). The binding sites are therefore independent. For each of the three fatty acids in the absence or in the presence of one or two of the others, the inside/outside distributions of the binding sites and the membrane transfer rate constants are elucidated by exchange efflux kinetics at 0°C from ghosts with and without enclosed albumin. Packed ghosts loaded with radioactive acids are injected rapidly into a large volume of vigorously stirred buffer with albumin. With a resolution time of about 1-sec serial filtered ghost-free aliquots are collected and counted. The analyses show that palmitic- and oleic acid sites of transport are entirely independent but do not exclude that palmitic- and/or oleic acid binding may diminish the arachidonic acid affinity a little. The diversity combined with specificity suggests that the transport sites for long chain fatty acids are protein-determined microdomains of phospholipids. Received: 26 June 1995/Revised: 11 October 1995     

Optokinetic speed control and estimation of travel distance in walking honeybees

Honeybees returning from foraging trips were video-filmed while they walked through a narrow transparent gangway to reach the hive entrance. On their way they were presented with black-and-white gratings viewed underneath as well as to both sides of the gangway. Bees could exit the gangway through one of two or three side exits installed at different distances from the gangway entrance. In one set of experiments, the substrate on which the bees walked was moved either in the bee's direction or against it. In another set of experiments, the substrate was stationary, but the pattern was moved in one or the other direction. The bee's walking speed (WS) was evaluated from the video tapes. When the substrate moved against, or the pattern in the bee's direction, in either case decreasing the speed of pattern flow (PFS), the bees increased WS, and, at the same time, they preferred the more distant exit. When the substrate moved in, or the pattern against the bee's direction, thus increasing PFS, WS decreased and the bees preferred the nearer exit. These results suggest that the speed of optic flow controls the speed of locomotion and might therefore also serve for assessing the distance travelled.Abbreviations PFS pattern-flow speed - WS walking speed Dedicated to Hans-Jochen Autrum, editor emeritus, for help in giving birth to this and many other papers over very many years, often with criticism but nevertheless with encouragement and sympathy.     

Integration time for short broad band clicks in echolocating FM-bats (Eptesicus fuscus)

Vespertilionid FM-bats (four Eptesicus fuscus and one Vespertilio murinus) were trained in an electronic phantom target simulator to detect synthetic echoes consisting of either one or two clicks. The threshold sound pressure for single clicks was around 47 dB peSPL for all five bats corresponding to a threshold energy of -95 dB re 1 Pa² * s. By varying the interclick interval, T, for double clicks it was shown that the threshold intensity was around — 3 dB relative to the threshold for single clicks at T up to 2.4 ms, indicating perfect power summation of both clicks. A threshold shift of -13.5 dB for a 1 ms train of 20 clicks (0.05 ms interclick interval) confirmed that the bats integrated the power of the stimuli. At T longer than around 2.5 ms the threshold for double clicks was the same as for single clicks. Thus, the bats performed like perfect energy detectors with an integration time of approximately 2.4 ms. This integration time is an order of magnitude shorter than that reported for bats listening passively for pure tones. In our setup the bats emitted sonar signals with durations of 2–3 ms. Hence, the results may indicate that while echolocating the bats integration time is adapted to the duration of the sonar emissions.Abbreviations AGC automatic gain control - FM frequency modulated - peSPL peak equivalent sound pressure level - rms root mean square - SD standard deviation - SE standard error of mean - T interclick interval     

Site of stimulation by mannosyl-P-dolichol of GlcNAc-lipid formation by microsomes of embryonic chick retina

Mannosyl-P-dolichol (man-P-dol) has been shown to stimulate the early reactions of the dolichol pathway, specifically, the biosynthesis of GlcNAc-P-P-dol and GlcNAc-GlcNAc-P-P-dol, and thus may play a regulatory role in glycoprotein biosynthesis. The site of action of man-P-dol has previously been suggested to be the GlcNAc-transferase concerned with the formation of the monoglucosaminyl derivative. Since the concentration of the chitobiosyl compound also increases as a result of the presence of man-P-dol, the immediate site of the activation was reexamined. The effect of man-P-dol on the formation of GlcNAc-GlcNAc-P-P-dol using GlcNAc-P-P-dol synthesizedin situ or added exogenously as the substrate was investigated. In addition, the distribution of radioactivity in the glucosaminyl constituents of the products under the stimulatory conditions was determined. The results of these studies supported the conclusion that the stimulation of GlcNAc-lipid synthesis by man-P-dol is due to the enhanced synthesis of GlcNAc-P-P-dol. It is not a result of the activation of the GlcNAc-transferase catalyzing the attachment of the second GlcNAc residue for the biosynthesis of the chitobiosyl derivative.Abbreviations GlcNAc-P-P-dol N-acetylglucosaminylpyrophosphoryldolichol - GlcNAc-GlcNAc-P-P-dol N-acetylglucosaminyl-N-acetylglucosaminylpyrophosphoryldolichol; - chito N-N-diacetylchitobiose - man-P-dol mannosylphosphoryldolichol - TX-100 triton X-100 - Tes 2-{[tris-(hydroxymethyl)-methyl]-amino}-ethanesulfonic acid     

On the relationship of thermodynamic parameters with the buried surface area in protein-ligand complex formation

Prediction of thermodynamic parameters of protein-protein and antigen-antibody complex formation from high resolution structural parameters has recently received much attention, since an understanding of the contributions of different fundamental processes like hydrophobic interactions, hydrogen bonding, salt bridge formation, solvent reorganization etc. to the overall thermodynamic parameters and their relations with the structural parameters would lead to rational drug design. Using the results of the dissolution of hydrocarbons and other model compounds the changes in heat capacity (C_p), enthalpy (H) and entropy (S) have been empirically correlated with the polar and apolar surface areas buried during the process of protein folding/unfolding and protein-ligand complex formation. In this regard, the polar and apolar surfaces removed from the solvent in a protein-ligand complex have been calculated from the experimentally observed values of changes in heat capacity (C_p) and enthalpy (H) for protein-ligand complexes for which accurate thermodynamic and high resolution structural data are available, and the results have been compared with the x-ray crystallographic observations. Analyses of the available results show poor correlation between the thermodynamic and structural parameters. Probable reasons for this discrepancy are mostly related with the reorganization of water accompanying the reaction which is indeed proven by the analyses of the energetics of the binding of the wheat germ agglutinin to oligosaccharides.     

Arabidopsis consensus intron sequences

We have analysed 998 Arabidopsis intron sequences in the EMBL database. All Arabidopsis introns to adhere to the :GU...AG: rule with the exception of 1% of introns with :GC at their 5 ends. Virtually all of the introns contained a putative branchpoint sequence (YUNAN) 18 to 60 nt upstream of the 3 splice site. Although a polypyrimidine tract was much less apparent than in vertebrate introns, the most common nucleotide in the region upstream of the 3 splice site was uridine. Consensus sequences for 5 and 3 splice sites and branchpoint sequences for Arabidopsis introns are presented.     

人脑和人血清胆碱酯酶三维结构的计算机模拟研究

本文以同源的电鳐胆碱酯酶（Ｔ．ＡＣｈＥ）的三维结构为模板,模拟预测了人脑和血清胆碱酯酶（Ｈ。ＡＣｈＥ和Ｈ．ＢｕＣｈＥ）的三维结构和活力中心的组成。指Ｔ．ＡＣｈＥ,Ｈ．ＡＣｈＥ和Ｈ．ＢｕＣｈＥ宁间结构差异,并讨论了ＡＣｈ和Ｈ。ＡＣｈＥ的对接（ｄｏｃｋｉｎｇ）。Ｈ。ＡＣｈＥ和Ｈ．ＢｕＣｈＥ三维结构的确定将为进一步深入研究它的中毒机理和合理药物设计提供靶子。     

籼稻232蜡质基因转录起始位点的鉴定

Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交分析和蜡质基因ｃＤＮＡ的序列分析表明水稻蜡质基因的转录本可能延伸到翻译起始密码子（ＡＴＧ）上游１２ｋｂ处。据此设计了２１Ｎｔ的寡核苷酸引物,并以籼稻２３２胚乳ＲＮＡ为模板,以引物延伸法确定籼稻２３２蜡质基因的转录起始点,籼稻蜡质基因的转录起始旁邻顺序ＣＴＣＡＣＣＡ与高等植物基因的转录起始点一致顺序ＣＴＣＡＴＣＡ仅相差１个碱基。通过顺序比较,对东乡野生稻蜡质基因中的转录起始位点的位置,以及对此两稻种中ＴＡＴＡ盒的可能顺序进行了讨论。     

乙肝病毒PreSl片段与乙肝表面抗原羧端的融合表达

我们利用聚合酶链反应法（ＰＣＲ）得到了编码乙肝病毒肝细胞受体结合位点ＰｒｅＳｌ（２１￣４７）的基因片段,并将它分别融合到Ｓ基因中相应于第１７５,１８８和２２３位氨基酸残基处。所得到的融合基因插入痘苗病毒表达载体ｐＧＪＰ－５后,在哺乳动物细胞ＣＶ－１中进行了暂时表达,对融合蛋白的表达、分泌和抗原性的研究表明,３种融合基因均能表达具有Ｓ和ＰｒｅＳｌ双重抗原性的融合蛋白,但融合位点对表达水平和分泌性质有     

病毒基因组启动子识别的人工神经网络方法

本文运用神经网络方法,并结合病毒基因分子生物学有关理论与统计事实,对病毒基因启动子区域进行了识别,文中选择了共35个基因组,作为研究对象．学习组选择了28个基因组,预测组选择了7个基因组,结果表明,将神经网络模型与病毒基因有关理论相结合,能够运用计算方法,以大量的可能启动子组合中排列出唯一的启动子区域．