Quantitative approaches to utilizing mutational analysis and disulfide crosslinking for modeling a transmembrane domain.

The transmembrane domain of chemoreceptor Trg from Escherichia coli contains four transmembrane segments in its native homodimer, two from each subunit. We had previously used mutational analysis and sulfhydryl cross-linking between introduced cysteines to obtain data relevant to the three-dimensional organization of this domain. In the current study we used Fourier analysis to assess these data quantitatively for periodicity along the sequences of the segments. The analyses provided a strong indication of alpha-helical periodicity in the first transmembrane segment and a substantial indication of that periodicity for the second segment. On this basis, we considered both segments as idealized alpha-helices and proceeded to model the transmembrane domain as a unit of four helices. For this modeling, we calculated helical crosslinking moments, parameters analogous to helical hydrophobic moments, as a quantitative way of condensing and utilizing a large body of crosslinking data. Crosslinking moments were used to define the relative separation and orientation of helical pairs, thus creating a quantitatively derived model for the transmembrane domain of Trg. Utilization of Fourier transforms to provide a quantitative indication of periodicity in data from analyses of transmembrane segments, in combination with helical crosslinking moments to position helical pairs should be useful in modeling other transmembrane domains.     

Characterization of a new four-chain coiled-coil: influence of chain length on stability.

Limited information is available on inherent stabilities of four-chain-coils. We have developed a model system to study this folding motif using synthetic peptides derived from sequences contained in the tetramerization domain of Lac repressor. These peptides are tetrameric as judged by both gel filtration and sedimentation equilibrium and the tetramers are fully helical as determined by CD. The four-chain coiled-coils are well folded as judged by the cooperativity of thermal unfolding and by the extent of dispersion in aliphatic chemical shifts seen in NMR spectra. In addition, we measured the chain length dependence of this four-chain coiled-coil. To this end, we developed a general procedure for nonlinear curve fitting of denaturation data in oligomeric systems. The dissociation constants for bundles that contain alpha-helical chains 21, 28, and 35 amino acids in length are 3.1 x 10(-12), 6.7 x 10(-23), and 1.0 x 10(-38) M3, respectively. This corresponds to tetramer stabilities (in terms of the peptide monomer concentration) of 180 microM, 51 nM, and 280 fM, respectively. Finally, we discuss the rules governing coiled-coil formation in light of the work presented here.     

降钙素和降钙素基因相关肽的选择性表达

降钙素和降钙素基因相关肽(CT/CGRP)由同一基因编码,该基因结构及其5＇端侧翼序列决定了它能够在甲状腺C细胞以及中枢和外周神经细胞生成不同的表达产物.这种选择表达调控决定多细胞生物的发育、性别分化和进化.如果表达失控将导致甲状腺髓样瘤(MTC)和骨质疏松症等疾病.文章对该基因的结构和选择性表达调控进行了综述.     

Identification and fine mapping of quantitative trait loci for the number of vascular bundle in maize stem

Studies that investigated the genetic basis of source and sink related traits have been widely conducted. However, the vascular system that links source and sink received much less attention. When maize was domesticated from its wild ancestor, teosinte, the external morphology has changed dramatically; however, less is known for the internal anatomy changes. In this study, using a large maize‐teosinte experimental population, we performed a high‐resolution quantitative trait locus (QTL) mapping for the number of vascular bundle in the uppermost internode of maize stem. The results showed that vascular bundle number is dominated by a large number of small‐effect QTLs, in which a total of 16 QTLs that jointly accounts for 52.2% of phenotypic variation were detected, with no single QTL explaining more than 6% of variation. Different from QTLs for typical domestication traits, QTLs for vascular bundle number might not be under directional selection following domestication. Using Near Isogenic Lines (NILs) developed from heterogeneous inbred family (HIF), we further validated the effect of one QTL qVb9‐2 on chromosome 9 and fine mapped the QTL to a 1.8‐Mb physical region. This study provides important insights for the genetic architecture of vascular bundle number in maize stem and sets basis for cloning of qVb9‐2.     

下丘脑内侧基底部去传入及内侧前脑束损毁对弓状核单位伤害性反应及电针反应的影响

在67只清醒麻痹的雄性大鼠身上,记录了157个弓状核(ARC)的单位放电。实验分四组:(1)正常对照组;(2)下丘脑内侧基底部(MBH)前去传入组;(3)MBH后去传入组;(4)内侧前脑束(MFB)损毁组。正常对照组ARC 单位的平均自发放电频率为2.03±0.3Hz(n=60),MBH前、后去传入组,ARC单位的平均自发放电频率比正常对照组显著减慢(P<0.01),说明从MBH前、后侧传入ARC的纤维对ARC单位有紧张性的兴奋作用。 在正常对照组,大多数ARC单位都能对隐神经伤害性刺激和电针起反应;但在MBH前去传入组,大多数ARC单位对隐神经伤害性刺激和电针不起反应。说明隐神经伤害性刺激和电针信息是从MBHM侧的神经通路传入ARC的。 在MFB损毁组,ARC单位对伤害性刺激的反应持续时间(2206.9±305.9ms,n=13)比正常对照组(1300.0±190.4ms,n=24)显著延长(P<0.05),说明MFB对ARC单位的伤害性反应有抑制性的调制作用。     

Protease-sensitive anchoring of microfilament bundles provides tracks for cytoplasmic streaming inVallisneria

Summary In mesophyll cells of the aquatic angiospermVallisneria gigantea Graebner, the endoplasm streams rotationally along the cell walls normal to the leaf surface in situ. Bundles of microfilaments anchored in the ectoplasm serve as tracks for the cytoplasmic streaming. In single mesophyll cells isolated by enzymatic digestion, hypertonic treatment induces abnormal streaming concomitant with plasmolysis, specifically at one or both of the shorter sides of an approximate rectangle. The disorderly arrangement of microfilaments in such cells has been confirmed by fluorescence microscopy of cells stained with FITC-phalloidin. While inhibitors of proteases added to the enzyme solution used for isolation of cells suppress the disturbance of rotational streaming, exogenously applied protease promotes it. The results suggest that bundles of microfilaments in the ectoplasm are stabilized by protease-sensitive factor(s) in the presence of the cell wall.     

Benzodiazepine receptors along the nephron: [3H]PK 11195 binding in rat tubules

Binding of [3H]PK 11195, an isoquinoline carboxamide derivative, was measured in microdissected tubule segments of rat nephron. High specific binding capacities (1.1-1.8 fmol X mm-1) were found in the thick ascending limb of the Henle's loop and in the collecting tubule, whereas specific binding could not be detected in the proximal tubule. In the medullary collecting tubule, the association and dissociation rate constants at 4 degrees C were k1 = 3.0 X 10(6) M-1 X min-1 and k-1 = 0.021 min -1; the ratio k-1/k1 = 7.0 nM was in agreement with the estimated equilibrium dissociation constant (Kd = 2.4 nM). [3H]PK 11195 binding sites from medullary ascending limb and medullary collecting tubule revealed the following sequence of specificity: PK 11195 = Ro 5-4864 much greater than clonazepam, indicating that tubule binding sites might be the peripheral benzodiazepine receptors of the rat kidney.     

玉米叶肉细胞和维管束鞘细胞中光合产物的分析

玉米苗照光后,叶肉细胞和维管束鞘细胞大量积累淀粉和可溶性糖(包括蔗糖),其中淀粉95％以上在维管束鞘细胞中。阻断光合产物输出时,两类细胞中蔗糖和淀粉积累都显著增加。离体维管束鞘细胞也能合成蔗糖。离体玉米叶内原生质体饲喂NaH~(14)CO_3并照光后,通常90％以上的~(14)C参入到有机酸和氨基酸中,3～10％参入糖和淀粉中。玉米叶肉原生质体具有直接利用CO_2合成碳水化合物的能力。     

广西野生蔗及其近缘植物营养器官内部结构的初步研究

广西野生蔗及其近缘植物营养器官内部结构基本相同。但割手密叶片下表皮具有明显的气孔窝斑茅等则无。斑茅茎节间的维管束分布较均匀,河八王茎中央没有维管束分布,割手密与蔗茅茎中维管束的分布。居于二者之间。