Modelling protein three-dimensional structure using tritium planigraphy

We propose the use of data on the topography of the label-accessible surface of a protein molecule obtained by the method of tritium planigraphy as a criterion for choosing the optimal intermediate arrangements of alpha-helices in globular proteins so as to model their three-dimensional structures. This approach has been used for modelling the three-dimensional structure of parvalbumin III from pike. The proposed model has been compared with high-resolution X-ray structural data for a related protein, paryvalbumin from carp. The possibilities and limitations of this approach are discussed.     

Megalodiscus temperatus: absorption and incorporation of tritiated tyrosine, thymidine, and adenosine

Autoradiographic and liquid scintillation counting studies indicate adults of Megalodiscus temperatus can absorb large amounts of thymidine and adenosine, but use little of the thymidine in in vitro exposures of up to 72 hr. Labeling of worms exposed to ³H-adenosine in periods as short as 3 hr was achieved especially over young eggs, testes, and the ovary. Large amounts of tyrosine are found in the free amino acid pool of this worm but little is incorporated into tissues for exposure periods of up to 24 hr. Ligation experiments and autoradiograms of frozen-dried worms show thymidine and adenosine can enter via the tegument. Adenosine added to the exposure medium did not inhibit absorption of tyrosine but did markedly reduce the uptake of ³H-thymidine in 8-hr in vitro exposures. Bacteria adhering to the glycocalyx of M. temperatus were identified as Escherichia coli, but their role in uptake phenomena is unknown. The association of thymidine with the lymph vessels indicates a circulatory or excretory role for this system in M. temperatus. Adenosine was considered the best compound of the three to use for labeling cells for studies on the reproductive system.     

Gorgoderina attenuata: uptake and incorporation of tyrosine, thymidine, and adenosine

In autoradiographic and scintillation counting studies, tritiated tyrosine, thymidine, and adenosine were found to be absorbed and incorporated by Gorgoderina attenuata in in vitro exposures of up to 12 hr. Tyrosine was incorporated into all tissues with the highest levels found in the intestinal epithelium. No evidence for selective incorporation of tyrosine into vitellaria was found. Thymidine was incorporated mainly into nuclei of gonial cells of the reproductive system and developing miracidia within eggs. Autoradiograms indicated assimilation of tyrosine and thymidine was highly variable among individual worms of a treatment group. Adenosine was absorbed and incorporated by worm tissues in large amounts within 3 hr. Variability of labeling on autoradiograms was not as great as with tyrosine and thymidine. Results of scintillation counting and grain counts of autoradiograms on ligated worms indicated all three compounds can enter the worm through the tegument. Adenosine was considered the best possible compound of the three to use for labeling cells for studies on the reproductive system.     

影响小麦粒重的农艺性状,生理指标的主成分分析

本研究对长江中游麦区11个小麦品种（系）的13个与粒重密切相关的农艺性状和生理指标进行了主成分分析。13个原始指标综合成为4个独立的新指标;主成分1,光合同化因子;主成分2,籽粒灌浆速率与构成因子;主成分3,籽粒灌浆持续期与产量构成因子;主成分4,籽粒灌浆参数综合因子。主成分值与粒重相关分析表明,主成分4及主成分2与粒重有极显著的正相关。通过逐步回归分析,建立了小麦粒重与主成分值之间的回归方程。以主成分值为指标的系数聚类,可将十一个品种分为三大类型。不同类型品种主成分值的大小和相互协调方式不同,直接导致品种（系）粒重不同。     

pH dependence of uptake hydrogenases in Azospirillum brasilense and A. amazonense

Abstract Methylene blue (MB)-dependent uptake hydrogenase activity of Azospirillum brasilense was at least 30% higher than O₂-dependent activity in this species, whereas for A. amazonense the reverse was true. Both MB-dependent and O₂-dependent uptake hydrogenase activities of A. brasilense showed maxima at pH 7.0 both in vivo and in vitro. For A. amazonense , both in vivo and in vitro, the MB-dependent uptake hydrogenase activity increased roughly linearly as pH values were increased from pH 6.25 to 7.75, whereas the O₂-dependent activity was almost unaffected by pH. The optimal pH for A. amazonense growth was broad and in the range of 5.4 to 6.5.     

Membrane transport in Schistosoma mansoni: transport of amino acids by adult males.

The mechanisms by which adult male Schistosoma mansoni transport amino acids have been investigated using radioactive amino acids during 2-min incubation times. The transport constants (K_t) for mediated uptake of glycine, proline, methionine, arginine, glutamate, and tryptophan were calculated to be 0.60-1.05, 1.67-1.98, 2.0, 0.10-0.35, 0.30-0.50, and 0.5-1.0 mM, respectively. Maximal velocities (V_max) were 5.5–7.5, 25, 6.4, 1.5-2.0, 2.5, and 3.0–6.0 μmoles absorbed/g worm protein/2 min, respectively. Cysteine is taken up solely by diffusion. Proline uptake is unique in that no significant diffusion component was found. The other amino acids studied were absorbed by diffusion as well as by specific transport systems. In the 2-min incubation periods employed glycine, proline, glutamate, and methionine were not significantly metabolized indicating that the uptake studies using these substrates reflect transport. Metabolism of the other amino acids used in these studies was not examined. The specificity of the transport systems was studied by testing the inhibitory effects of various amino acids on the uptake of each of the amino acids studied. The results suggest the presence of at least five transport systems. There is a highly specific transport locus for proline, and one for acidic amino acids. There are probably at least two transport systems, each of broad and overlapping specificity, for most of the neutral amino acids. Basic amino acids also appear to be taken up by complex transport systems, at least one of which overlaps with the neutral sites. The results are discussed with respect to the nutrition of the parasite and the host-parasite relationship.     

Regeneration of asymmetric somatic hybrid plants from the fusion of two types of wheat with Russian wildrye

Two types of protoplasts of wheat (Triticum aestivum L. cv. Jinan 177) were used in fusion experiments—cha9, with a high division frequency, and 176, with a high regeneration frequency. The fusion combination of either cha9 or 176 protoplasts with Russian wildrye protoplasts failed to produce regenerated calli. When a mixture of cha9 and 176 protoplasts were fused with those of Russian wildrye, 14 fusion-derived calli were produced, of which seven differentiated into green plants and two differentiated into albinos. The morphology of all hybrid plants strongly resembled that of the parental wheat type. The hybrid nature of the cell lines was confirmed by cytological, isozyme, random amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) analyses. GISH analysis revealed that only chromosome fragments of Russian wildrye were transferred to the wheat chromosomes of hybrid calli and plants. Simple sequence repeat (SSR) analysis of the chloroplast genome of the hybrids with seven pairs of wheat-specific chloroplast microsatellite primers indicated that all of the cell lines had band patterns identical to wheat. Our results show that highly asymmetric somatic hybrid calli and plants can be produced via symmetric fusion in a triparental fusion system. The dominant effect of two wheat cell lines on the exclusion of Russian wildrye chromosomes is discussed.Abbreviations GISH Genome in situ hybridization - RAPD Random amplified polymorphic DNA - SCF Small chromosome fragment - SSR Simple sequence repeat     

Ribosomal protein S1 induces a conformational change of the 30S ribosomal subunit

A comparative study of the 30S ribosomal subunit in the complex with protein S1 and the subunit depleted of this protein has been carried out by the hot tritium bombardment method. Differences in exposure of some ribosomal proteins within the 30S subunit depleted of S1 and within the 30S–S1 complex were found. It was concluded that protein S1 binds in the region of the neck of the 30S ribosomal subunit inducing a conformational change of its structure.     

Application of tritium high resolution NMR spectroscopy to analysis of tritium-labelled amino acids and peptides

Summary A method has been developed for the qualitative and quantitative analysis of complex isotopic mixtures of tritium-labelled amino acids and peptides by using high resolution³H NMR spectroscopy at 266.8 MHz. Determined were tritium distribution in alanine, glycine, tryptophan and 4-hydroxyproline amino acids, as well as in glycine and valine residues of peptides. Approaches have been worked out for the determination of spin coupling constants and isotope chemical shifts for the strongly coupled nonequivalent atoms of the methylene groups.