Crystallization and preliminary diffraction studies of malate dehydrogenase from Streptomyces aureofaciens

Purified malate dehydrogenase (MDH) of Streptomyces aureofaciens was crystallized either in the absence or in the presence of NADH or NADPH coenzymes by hanging-drop vapour-diffusion method. An X-ray study has shown, that MDH crystals belong to space group C222(1) with unit-cell parameters a = 53.2 A, b = 104.6 A, c = 520.0 A, alpha = beta = gamma = 90( degrees ), MDH-NADH crystals to space group C2 with unit-cell parameters a = 51.5 A, b = 51.5 A, c = 256 A, alpha = beta = gamma = 90( degrees ), and MDH-NADPH crystals to space group C222(1) with unit-cell parameters a = 72, A b = 72 A, c = 520 A, alpha = beta = gamma = 90( degrees ). The crystal of native MDH diffracted to 2.1 A resolution.     

国产十四种苋属植物的染色体数目

报道了国产14种苋属植物的染色体数目。部分种的染色体数目为2n=34,即反枝苋Amaranthus retroflexus,刺苋A．spinosus,红苋A．cruentus,腋花苋A．roxburghianus,合被苋A．polygonoides,皱果苋A． viridis,凹头苋A．lividus,苋A．tricolor。其他种的染色体数目为2n=32,即尾穗苋A．caudatus,绿穗苋A． hybridus,千穗谷A．hypochendriacus,繁穗苋A．paniculatus,北美苋A．blitoides,白苋A．albus。其中腋花 苋的染色体数目为首次报道。该属染色体基数为x=16,17。两种染色体基数在苋属2个组(sect．Ama- ranthus和sect．Blitopsis)中均存在。由于苋属植物染色体大多为小型染色体,因此对苋属植物目前尚不 能进行详尽的核型分析。     

黄芪属六种植物的核型多样性

采用常规压片法,对豆科黄芪属6种植物制备染色体标本进行核型分析。结果表明:体细胞中期染色体数目分别为:沙打旺、斜茎黄芪、达乌里黄芪2n=16,均为二倍体;草木樨状黄芪2n=32,为四倍体;紫云英、鹰嘴紫云英则呈现多数目性,紫云英染色体数变动范围为55～65,64条稍多,鹰嘴紫云英染色体数变动范围51～65,62条稍多,均为混倍体。核型公式分别为:沙打旺2n=2x=16=12m+4sm;斜茎黄芪2n=2x=16=10m+6sm;达乌里黄芪2n=2x=16=16m;草木樨状黄芪2n=4x=32=32m;紫云英2n=64=62m+2sm;鹰嘴紫云英2n=62=12M+50m(2SAT)。染色体核型呈现多样性。     

国产十五种翠雀族植物的核型研究

本文报道了国产乌头属Aconitum L.12种和翠雀属Delphinium L．3种植物的染色体数目和核型。花葶乌头A．scaposum var．scaposum和聚叶花葶乌头A．scaposum var．vaginatum的核型公式为2n=2m+6sm+8st;两色乌头A．alboviolaceum为2n＝16＝2m+6sm(2sat)十8st;高乌头A．sinomontanum var．sinomontanum为2n＝16＝4m+4sm+8st,而狭盔高乌头A．sinomontanum var．angustius为2n＝32＝6m+6sm+20st(1sat);褐紫乌头A.brunneum为2n＝16＝2m(1sat)+14sm 松潘乌头A．sungpanense为2n＝16＝6m(2sat)+10sm(2sat);弯枝乌头A．arcuatum为2n=16=4m+12sm;乌头A．carmichaeli为2n=32＝10m+22sm;北乌头A．kusnezoffii为2n＝32＝10m十22sm;鸭绿乌头A.jaluense为2n＝32=10m+22st;伏毛铁棒锤A．flavum为2n＝16＝2m+14sm; 铁棒锤A．pendulum为2n=16=4m+12sm;缩梗乌头A．sessiliflorum为2n＝16＝2m+10sm+4st;展毛翠雀花D．kamaoense var．glabrescens 为2n=16＝2m+6sm+8st;蓝翠雀花D．caeruleum为2n=16=2m+6sm+8st;多枝翠雀花D．maximowiczii为2n=16=2m+6sm+8st。其中花葶乌头、狭盔高乌头、褐紫乌头、松潘乌头、鸭绿乌头、伏毛铁棒锤、缩梗乌头、展毛翠雀花、蓝翠雀花、多枝翠雀花的染色体数目和核型为首次报道。     

东北蒿属艾组12种植物核型研究

报道东北蒿属艾组12种植物的染色体数及核型资料,结果如下：宽叶山蒿（Artemisia stolonifera (Maixm.)Komar.)2n=4x=36=28m 4sm(4SAT) 4st(4SAT);野艾蒿（A.lavandulaefolia DC)2n=2x=50=40m 4sm 6st(6SAT); 矮蒿（A.lancea Van.)2n=2x=16=10m 4sm 2st;蒙古蒿（A.mongolica (fisch.ex Bess.)Nakai]2n=2x=16=14m 2st;红足蒿（A.rubripes Nakai)2n=2x=16=14m 2st;歧茎蒿（A.igniaria Maixm.)2n=2x=34=28m 2sm 4st(4SAT);柳叶蒿（A.integrifolia Linn.)2n=4x=36=20m 12sm 4st;线叶蒿（A.subula 4st(4SAT);柳叶蒿（A.integrifolia Linn.)2n＝4x=36=20m 12sm 4st;线叶蒿（A.subulata Nakai)2n=2x=16=14m 2st;高岭蒿（A.brachyphylla Kitam.)2n=2x=18=14m 2sm 2st;林文稿[A.uiridissima(Komar.)Pamp.]2n=2x=18=14m 4sm;奄癌（A.keiskeana Miq.)2n=2x=18=16m 2sm;阴地蒿（A.sylvatica Maxim.)2n=2x=16=14m 2sm,核型对称性野艾蒿（A.lavandulaefolia)为2B型,其余均为2A型,依据核型资料对个别种的演化 分类进行了初步探讨。     

Crystallization of murine major histocompatibility complex class I H-2Kb with single peptides.

X-ray quality crystals of a soluble murine class I H-2Kb molecule complexed with three different peptide antigens were grown in several forms by streak seeding and macroseeding methods. Co-crystals with VSV-8 (RGYVYGQL), OVA-8 (SIINFEKL) and SEV-9 (FAPGNYPAL) peptides were grown either from NaH2PO4/HPO4 or from polyethylene glycol 4000 within the pH range 5.0 to 7.5, with the use of 4-methyl-2-pentane diol (MPD) as an additive. The VSV-8 crystals grew in space groups P1, with cell dimensions a = 63.1 A, b = 69.1 A, c = 72.0 A, alpha = 89.9 degrees, beta = 77.1 degrees, gamma = 123.3 degrees and P2(1)2(1)2, with a = 138.1 A, b = 88.6 A, c = 45.7 A, and diffract to 2.9 and 2.3 A, respectively. Crystals of the SEV-9 complex grew from similar crystallization conditions to those of the orthorhombic VSV-8 complex with similar cell parameters and diffract to at least 2.5 A resolution. Crystals of the OVA-8 complex were obtained from either phosphate (space group C2, a = 118.7 A, b = 61.6 A, c = 85.3 A, beta = 108.4 degrees) or polyethylene glycol (space group P1, a = 64.5 A, b = 71.0 A, c = 66.3 A, alpha = 89.7 degrees, beta = 95.7 degrees, gamma = 123.3 degrees) and diffract to 3 A resolution. The crystallization procedures used here significantly increased the rate and production of X-ray quality crystals.     

国产6种黄耆属植物的核型研究

报道了国产黄耆属(Astragalus)6种植物的染色体数目和核型。结果表明,这6种黄耆属植物均为二倍体,其核型公式分别为：甘青黄耆(A．tangutlcus),2n=16=6m 8sm 2t,“3A”核型;悬垂黄耆(A．dependens),2n=16=10m 6sm,“2A”核型;四川黄耆(A．sutchuenensis),2n=16=8m 8sm,“2A”核型;萨雷古拉黄耆(A.pavlovlanus),2n=16=12m 4sm,“1A”核型;喜石黄耆(A.petraeus),2n=16=14m 2sm,“1A”核型;拟糙叶黄耆(A.pseudoscaberrimus),2n=16=14m 2sm,“1A”核型。这6种黄耆属植物的染色体数目和核型均为首次报道。     

中国西北地区11种黄耆属植物的细胞学研究

对分布于中国西北地区的11种黄耆属植物进行了细胞学研究,其中9种作了核型分析。结果表明,杯萼黄耆（Astragalus cupulicalycinus）为六倍体（2n=48）,酒泉黄耆（A.jiuquanensis）具有2种细胞型：2n=32的四倍体和2n=48的六倍体,其余种类均为二倍体（2n=16)。9个种的核型公式分别为：木黄耆（A.arbuscula）,2n=16=14 2sm,“1A”核型;胀萼黄耆（A.ellipsoideus）,2n=16=10m 6sm,“2A”核型;粗毛黄耆（A.scabrisetus),2n=16=14m 2sm,“1A”核型;秦岭黄耆（A.henryi),2n=16=8m 8sm(2SAT),“2A”核型;哈巴河黄耆（A.habaheensis）,2n=16=10m 6sm(2SAT),“2A”核型;变异黄耆（A.variabilis）,2n=16=4m 10sm(2SAT) 2t,“2A”核型;喜沙黄耆（A.ammodytes),2n=16=6m 10sm(2SAT),“2A”核型;密花黄耆（A.densiflorus),2n=16=6m(2SAT) 10sm,“2A”核型;茧荚黄耆（A.lehmannianus),2n=16=14m(2SAT) 2st,“2A”核型。所有研究种中,仅变异黄耆的染色体数目有过报道,其余种的染色体数目及核型均为首次报道。     

Molecular size and symmetry of the bacterioferritin of Escherichia coli. X-ray crystallographic characterization of four crystal forms

X-ray crystallographic data from four crystal forms of Escherichia coli bacterioferritin show that the molecule has a diameter in the range 119 to 128 A. Molecules are composed of 24 subunits arranged in 432 symmetry. In both size and symmetry the molecule resembles ferritin from eukaryotes. The four crystal forms are monoclinic, space group P2(1) with unit cell dimensions a = 118.7 A, b = 211.6 A, c = 123.3 A and beta = 119.1 degrees; orthorhombic, C222(1), a = 128.7 A, b = 197.1 A, c = 202.8 A; tetragonal, P4(2)2(1)2, a = b = 210.6 A, c = 145.0 A and cubic, I432, a = 146.9 A.     

Preparation and X-ray characterization of four new crystal forms of jacalin, a lectin from Artocarpus integrifolia

Four new crystal forms of the anti-T lectin from jackfruit (Artocarpus integrifolia) have been prepared and characterized. Three of them, two monoclinic (P21, a = 59.4 A, b = 83.3 A, c = 63.5 A, beta = 107.7 degrees; C2, a = 106.1 A, b = 53.9 A, c = 128.0 A, beta = 95.0 A) and one orthorhombic (C222(1), a = 98.1 A, b = 67.3 A, c = 95.1 A) were grown with 2-methylpentan-2,4-diol (MPD) as the precipitant while the fourth, an hexagonal form (P6(1)22, a = b = 129.6 A, c = 157.9 A), was obtained in the presence of methyl-alpha-D-galactopyranoside with polyethylene glycol 4000 as the precipitant. The reported relative molecular mass (Mr) of the lectin was found to be inconsistent with the solvent content of the crystals estimated using measured densities. The Mr was redetermined using size-exclusion chromatography in the presence of methyl-alpha-D-galactopyranoside and Ferguson-plot analysis of mobilities in polyacrylamide gel electrophoresis. The redetermined Mr (66,000) is consistent with the measured crystal densities. The orthorhombic and the hexagonal forms, which have one half molecule and one molecule, respectively, in the asymmetric unit, are suitable for high-resolution X-ray analysis.     

Crystallization and preliminary crystallographic data of the Fab fragment of an anti-phenylalanine hydroxylase monoclonal antibody

Two crystal habits, one rod shaped and the other square prismatic, of the Fab fragment of a monoclonal anti-phenylalanine hydroxylase antibody have been grown using the method of vapour phase diffusion against polyethylene glycol 6000. The square prisms diffract to better than 2.8 A, belong to the space group P1 and have unit cell parameters a = 41.8 A, b = 50.3 A, c = 114.7 A, alpha = 97.6 degrees, beta = 91.7 degrees, gamma = 91.0 degrees, while the rod-shaped crystals belong to the space group P212121, have unit cell parameters a = 105.6 A, b = 119.8 A, c = 82.2 A and diffract to 3.5 A resolution.     

Crystallization of nitrite reductase from Achromobacter cycloclastes

Crystals of a green copper-containing nitrite reductase from Achromobacter cycloclastes, which diffract to high resolution, belong to the cubic space group P213, with a = b = c = 98.4 A. Crystals of a nitrite reductase from Alcaligenes faecalis S-6 have been made, and belong to space group P212121, with a = 77.3 A, b = 94.6 A and c = 141 A. Crystals of the blue copper protein from Ac. cycloclastes have also been obtained: these belong to space group P21212, with cell dimensions a = 34.9 A, b = 91.1 A and c = 36.7 A (1 A = 0.1 nm).     

Chromosome Numbers of 10 Species and 3 Varieties in Actinidia Lindl.

Somatic chromosome numbers of 10 species and 3 varieties in Actinidia Lindl. are reported for the first time, except A. polygama (Sieb. et Zucc.) Maxim. Two ploidy levels, diploid and tetraploid, are found both in A. polygama and A. kolomikta (Maxim. et Rupr.) Maxim. Chromosome numbers are listed as follows: A. kolomikta (Maxim. et Rupr.) Maxim. 2n=58, 116, A. polygama (Sieb. et Zucc. Maxim. 2n=58, 116, A. macrosperma Liang 2n=116, A. callosa var. henryi Maxim. 2n=116, A. callosa var. discoler Liang 2n =116, A. cylindrica Liang 2n=58, A. chrysantha Liang 2n=116, A. sabiaefolia Dunn 2n= 58, A. melliana Hand.-Mazz. 2n=58, A. hemsleyana Dunn 2n=58, A. fulvicoma var. lanata (Hemsl.) Liang 2n=58, A. latifolia (Gardn. et Champ.) Merr. 2n=58, and A. lanceolata Dunn 2n=58.     

Crystallization and preliminary crystallographic studies of an antitumour lectin from the edible mushroom Agrocybe aegerita

An antitumour lectin named AAL has been purified from the fruiting body of edible mushroom Agrocybe aegerita. In addition to having a distinct bioactivity, AAL shows strong inhibition effects on human and mouse tumour cells. It has been shown that AAL exerts its antitumour effects via apoptosis-induction. AAL and AAL-lactose complex have been crystallized and their diffraction data were collected with resolution of 2.6 A and 3.0 A, respectively. Both crystals belong to space group P 6122 with unit cell parameters a = 123.98 A, b = 123.98 A, c = 56.86 A, alpha= beta=90 degrees, gamma = 120 degrees and a = 123.69 A, b = 123.69 A, c = 56.64 A, alpha = beta = 90 degrees, gamma = 120 degrees, respectively.     

国产８种蜘蛛抱属植物的核型研究

首次报道了８种蜘蛛抱属植物的核型,其中６种的染色体数目为首次报道,结果如下：峨边蜘蛛抱蛋Ａ.ebianensis,2n=2x=36=18m 2sm(2sat) 16st;盈江蜘蛛抱蛋Ａ．yingjiangensis,2n=2x=36=14m 6sm(2sat) 16st;海南蜘蛛抱蛋Ａ．hainensis,2n=2x=36=20m(2sat) 14st 2t;石山蜘蛛抱蛋Ａ.saxicola,2n=2x=36=16m 4sm(2sat) 16st;要蜘蛛抱蛋白Ａ.muricata,2n=2x=36=18m 2sm(2sat) 16st);啮边蛛抱蛋Ａ.marginella,2n=2x=38=22m 4sm(2sat) 12st;西林蜘蛛抱蛋Ａ.xillinensis,2n=4x=76=48m(4sat) 2sm 26st;十字蜘蛛抱蛋A.cruciformis,2n=4x=76=46m(4sat) 12sm 18st。核型类型都为２Ｃ型。首次在中国发现了Ａ.cruciformis和A.xilinensis的野生四倍体。根据外部形态性及已有的３８种植物的核型资料分析,认为该属染色体的原始基数可能为x=18,核型向对称性增强的方向演化,其主要表现在中部着丝粒染色体数目的增多,这种演化趋似与其花部结构的进化密切相关。     

Crystallization and preliminary diffraction studies of recombinant human granulocyte-stimulating factor (KW2228).

Human granulocyte colony-stimulating factor (hG-CSF) specifically stimulates proliferation of neutrophils. Two crystal forms of a mutant of hG-CSF expressed in Escherichia coli have been obtained using the hanging drop vapour diffusion method. One form is triclinic, space group P1, with cell dimensions a = 37.3 A, b = 46.4 A, c = 47.7 A, alpha = 105.5 degrees, beta = 98.0 degrees and gamma = 109.4 degrees. The other is monoclinic, space group C2, with cell dimensions a = 82.0 A, b = 49.2 A, c = 49.4 A and beta = 113.9 degrees. Both crystal forms diffract beyond 2.0 A and are suitable for X-ray analysis.     

Chromosome numbers of AlchemUla series Vulgares from NE Anatolia (Turkey)—a preliminary report

This paper presents the chromosome numbers of nine species from the section AlchemUla , subsection Heliodrosium , series Vulgares. The studies were carried out on plants from the Northeast Anatolia (Turkey). The following chromosome numbers have been detected; A. valdehirsuta 2n= 96–106, A. monticola 2n= 101–110, A. stevenii 2n= 98–106, A. crinita 2n= 102–108, A. compactilis 2n= 98, A. heterophylla 2n= 100–105, A. minusculiflora 2n= 87–97, A. pseudocartalinica 2n= 98–108, A. straminea 2n= 80–91. The species of genus AlchemUla studied from Northeast Anatolia are proven to be polyploids like the majority of species from the section AlchemUla investigated from various geographic areas.     

Crystallization and preliminary diffraction analysis of cholesterol esterase from Candida cylindracea

Cholesterol esterase (EC 3.1.1.13) from the microorganism Candida cylindracea has been crystallized in two forms. Crystals, typically 0.30 x 0.15 x 0.10 mm in size, diffract rotating anode generated x-rays to beyond 3 A are suitable for data collection for an x-ray crystallographic investigation. A monoclinic crystal form in the space group P2(1) was found to have cell dimensions of a = 122.9 A, b = 101.0 A, c = 95.2 A and beta = 108.3 degrees. The asymmetric unit of the cell contains two dimers of 129 kDa each. A second crystal form, in the triclinic space group P1, has cell dimensions of a = 58.6 A, b = 88.7 A, c = 58.6 A, alpha = 93.3 degrees, beta = 113.8 degrees and gamma = 96.0 degrees, and has one dimer per asymmetric unit.     

Association of cytochrome P450 genes polymorphisms (CYP1A1 and CYP1A2) with the development of chronic obstructive pulmonary disease in Bashkortostan

To assess the role that polymorphisms of cytochrome P450 genes play in genetic predisposition to chronic obstructive pulmonary disease (COPD), the allele and genotype distributions of CYPIA1 (2455 A/G, 3801T/C) and CYP1A2 (-2464T/delT, -163C/A) genes were studied in Tatar and Russian COPD patients and in cases of healthy individuals (Russian, Tatar and Bashkir), residents of Bashkortostan. It was shown that the CYP1A1 and CYP1A2 genes haplotypes frequency distribution patterns do not differed between Tatars and Russians ethnic groups (chi2 = 0.973, df = 3, p = 1.00 and chi2 = 1.546, df = 3, p = 0.92, respectively). Analysis of the the CYP1A1 and CYP1A2 genes haplotypes revealed statistically significant differences in the haplotypes frequency distributions between Bashkirs versus Russians and Tatars (chi2 = 12.328, df= 3,p = 0.008; chi2 = 9.218, df=3, p = 0.034, respectively for CYP1A1 gene and (chi2 = 18.779, df=3, p = 0.0001, chi = 14.326, df=3, p = 0.003, respectively for CYP1A2 gene). The (-2467)delT allele and CYP1A2*1D haplotype of CYPIA2 gene was associated with higher risk of COPD in Tatar ethnic group (OR = 1.83, 95% CI 1.24-2.71, chi2 = 9.48, p = 0.003 and chi2 = 9.733, p = 0.0027, Pcor = 0.008; OR = 3.908, 95% CI 1.56-10.19, respectively). On the other hand the CYP1A2*1A haplotype had protective effect (chi2 = 6.319, p = 0.0127, Pcor = 0.038; OR = 0.6012, 95% CI 0.402-0.898). But at the same time we did not find any differences in the genotypes and haplotypes frequency distributions of the CYP1A2 gene within the patients and healthy groups in Russian ethnic group. We also did not find any association of CYP1A1 gene with COPD in ethnic groups of Bashkortostan.     

Preliminary crystallographic study of bluetongue virus capsid protein, VP7.

Bluetongue virus serotype 10 (BTV-10) VP7, expressed by insect cells infected with the recombinant baculovirus, has been purified and crystallized. Two crystal forms suitable for X-ray analysis have been obtained. Type I crystals belong to space group P6(3)22 with a = b = 95.2 A, c = 181.0 A, alpha = beta = 90 degrees gamma = 120.0 degrees, and contain a single subunit in the crystallographic asymmetric unit. They diffract to dmin = 3.0 A. Type II crystals belong to space group P2(1) with a = 69.4 A, b = 97.1 A, c = 71.4 A, beta = 109.0 degrees, and contain a trimer in the crystallographic asymmetric unit. They diffract to dmin = 2.1 A. These results, together with solution studies, show that the molecule is a trimer.