Production, crystallization, and preliminary X-ray analysis of rabbit skeletal muscle troponin complex consisting of troponin C and fragment (1-47) of troponin I.

Troponin is a ternary protein complex consisting of subunits TnC. TnI, and TnT, and plays a key role in calcium regulation of the skeletal and cardiac muscle contraction. In the present study, a partial complex (CI47) was prepared from Escherichia coli-expressed rabbit skeletal muscle TnC and fragment 1-47 of TnI, which is obtained by chemical cleavage of an E. coli-expressed mutant of rabbit skeletal muscle TnI. Within the ternary troponin complex, CI47 is thought to form a core that is resistant to proteolytic digestion, and the interaction within CI47 likely maintains the integrity of the troponin complex. Complex CI47 was crystallized in the presence of sodium citrate. The addition of trehalose improved the diffraction pattern of the crystals substantially. The crystal lattice belongs to the space group P3(1)(2)21, with unit cell dimensions a = b = 48.2 A, c = 162 A. The asymmetric unit presumably contains one CI47 complex. Soaking with p-chloromercuribenzenesulfonate (PCMBS) resulted in loss of isomorphism, but enhanced the quality of the crystals. The crystals diffracted up to 2.3 A resolution, with completeness of 91% and R(merge) = 6.4%. The crystals of PCMBS-derivative should be suitable for X-ray studies using the multiple-wavelength anomalous diffraction technique. This is the first step for elucidating the structure of the full troponin complex.     

Direct micro-sequence analysis of peptides from Escherichia coli ribosomal proteins S11, L9 and L29 after separation by reversed phase chromatography

Tryptic peptides of the ribosomal proteins S11, L9 and L29 were separated by reversed phase chromatography under conditions which enabled direct micro-sequencing with the 4-(dimethylamino)azobenzene-4'-isothiocyanate/phenylisothiocyanate double coupling method [Chang, Brauer, Wittmann-Liebold (1978) FEBS Lett. 93, 205-214]. The peptides were separated on a RP-18 column employing volatile buffers at pH 2.0, 4.1 and 7.8. Depending on the different chromatographic behaviour of the peptide mixture, the elution gradient was optimised for each hydrolysate using 20 micrograms of the hydrolysed protein. Preparative separations were made with 150-250 micrograms. At least 80% of the peptides could be isolated by these techniques and used for direct micro-sequencing without further purification or desalting. The results show that the high-performance liquid chromatographic method employed allows easy isolation and sequencing with minute amounts of peptides.     

Conversion of sterigmatocystin to aflatoxin B 1 by Aspergillus parasiticus

¹⁴C-Sterigmatocystin isolated from cultures of $\text{Aspergillus}$$\text{versicolor}$ supplemented with (1-¹⁴C)acetate was shown to be efficiently converted to aflatoxin B₁ by the resting mycelium of $\text{A. parasiticus}$. The experimental results may indicate a biosynthetic pathway leading from 5-hydroxysterigmatocystin to sterigmatocystin and then to aflatoxin B₁.     

Ce~（3＋）与G－肌动蛋白结合引起的构象变化及对聚合的影响

用Ａｅｄａｎｓ标记肌动蛋白单体Ｇ－Ａｃｔｉｎ上Ｃｙｓ３７４残基作为探针,研究了稀土离子Ｃｅ~（３＋）与Ｇ－Ａｃｔｉｎ的结合及引起的微构象变化。Ｃｅ~（３＋）在低浓度（Ｃｅ~（３＋）／Ａｃｔｉｎ摩尔比＜１）和Ｃａ~（２＋）竞争Ｇ－Ａｃｔｉｎ上二价离子的高亲合位点。Ｃｅ~（３＋）取代Ｃａ~（２＋）引起Ａｅｄａｎｓ荧光强度增强与Ｍｇ~（２＋）取代Ｃａ~（２＋）的结果相同。Ｃｅ~（３＋）／Ａｃｔｉｎ＞ｌ则导致Ａｅｄａｎｓ荧光强度下降。说明Ｃｅ~（３＋）在高低两种浓度条件下结合的位点及对Ｃｖｓ３７４的微构象的影响不同。时间分辩测得的Ａｅｄａｎｓ荧光寿命也支持这一结论。ＣＤ谱结果表明Ｃｅ~（３＋）／Ａｃｔｉｎ＜０．４,Ａｃｔｉｎ的二级结构增加,大于０．４又导致其失去。Ｃｅ~（３＋）－Ａｃｔｉｎ在有／无游离ＡＴＰ时用聚合液诱导的聚合结果表明,无游离ＡＴＰ时,极低浓度Ｃｅ~（３＋）促进聚合,高浓度虽有促进但有所减弱;有游离ＡＴＰ时,Ｃｅ~（３＋）／Ａｃｔｉｎ在实验范围内促进聚合。     

中药固真方对一些与细胞增殖有关基因表达的影响

 中药固真方对一些与细胞增殖有关基因表达的影响姚明忠,顾文聪,丁卫,韩志芬,杜国光（上海中医药大学生物化学教研室,上海２０００３２）（北京医科大学生物化学与分子生物学系,北京１０００８３）中药固真方（ＶＲＦ）具有补肾益精、延缓衰老的作用［１］．能提高成...     

EB病毒BNLF-1基因的分子生物学研究进展

位于EB病毒基因组U₅-TR区内的BNLF-1基因,其转译产物为潜伏膜蛋白(latent membrane protein1, LMP-1),由于LMP-1可以导致细胞转化并在EB病毒致癌过程中具有重要作用,因而成为近年来EB病毒分子生物学及相关肿瘤如人鼻咽癌、伯基特淋巴瘤、何杰金氏病等疾病病因发病学研究的热点,并取得了一批有重要意义的成果,文章从BNLF-1的基因结构及表达调控, LMP蛋白的结构及生化功能, LMP-1的生物学功能和LMP-1研究进行评述.     

Serum lipoproteins and albumin in the lecithin: Cholesterol acyltransferase reaction

The unique features of pig ovarian follicular fluids, i.e., presence of high density lipoprotein (HDL) only and lecithin: cholesterol acyltransferase (EC 2.3.1.43; LCAT) activity, provides a good model to study the effect of serum lipoproteins and serum albumin on the LCAT reaction. $\text{In}\text{vitro}$ cholesterol esterification is enhanced when very low density lipoprotein (VLDL) and low density lipoprotein (LDL) fractions are added, but is inhibited when one or the other of these lipoproteins is absent. High concentrations of HDL₂ result in decreased activation which can be compensated for by the addition of the VLDL-LDL mixture. These findings suggest that the rate of cholesterol esterification in ovarian follicular fluid may be enhanced by providing the exogenous VLDL and LDL as the recipients of HDL-cholesteryl ester. The inhibition of LCAT activity caused by free fatty acid and lysophosphatidylcholine can be partially reversed by the addition of serum albumin, suggesting that serum albumin may regulate the LCAT reaction.     

天山林区六种灌木生物量的建模及其器官分配的适应性

灌木全株生物量估算模型的构建仍存在一定困难,对灌木生物量在器官分配上所体现的适应性研究也不够充分。以天山林区6种常见灌木为研究对象,在天山的东段、中段、西段林区分别设置样地进行群落调查,由此以全株收获法取得6种常见灌木若干标准株的全株、根、枝、叶及各径级根的生物量,将D~2H(地径平方与高度的乘积)与V(冠幅面积与高度的乘积)分别选为估测模型的自变量,通过回归分析法建立了各种灌木全株生物量的最优估算模型,然后比较了此6种灌木全株生物量在营养器官上分配差异以及根系生物量在径级上的分配差异。结果表明:(1)天山林区6种常见灌木中,小檗(Berberis heteropoda Schrenk)、忍冬(Lonicera hispida Pall.ex Roem.et Schuet.)、栒子(Cotoneaster melanocarpus Lodd.)的全株生物量约为8.48—9.01 kg,蔷薇(Rosa spinosissima L.)、绣线菊(Spiraea hypericifolia L.)、方枝柏(Juniperus pseudosabina Fisch.et Mey.)的全株生物量约为2.71—3.20 kg;(2)蔷薇、绣线菊、栒子的全株生物量最优估测模型是以V为自变量的函数,小檗、忍冬、方枝柏的全株生物量最优估测模型是以D~2H为自变量的函数,各模型R~2值均在0.850以上,且在P0.05水平上达到显著,模型模拟结果达到了较高的准确度;(3)6种灌木全株生物量在根、枝上的分配比重差异不显著,仅在叶上的分配比重有差异(P0.05);根系生物量在径级上的分配均呈现随根系径级下降而减少的规律,6种灌木在径级大于2 mm根上的分配比重存在差异(P0.05,径级大于20 mm根为P0.01水平);(4)6种灌木全株生物量在营养器官上的分配差异以及根系生物量在径级上的分配差异均体现了各物种对其生境选择的适应策略。