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991.
992.
脑型一氧化氮合成酶的钙调蛋白结合区的表达及活性鉴定 总被引:1,自引:0,他引:1
用PCR法克隆出nNOS的CaM结合区基因(nNOS 2455~2988bp),并在大肠杆菌中进行了高效表达。经金属离子螯合亲和层析得到纯度为90%以上的重组蛋白.分子量为22kDa,CaM Oveday assay证实该蛋白具有CaM的结合活性。由于所表达的重组蛋白既具有序列特异性又具有CaM的结合活性.因此。可将它作为筛选nNOS特异性抑制肽的靶蛋白,亦可用于特异性抗体的制备。 相似文献
993.
A recessive ryanodine receptor 1 mutation in a CCD patient increases channel activity 总被引:1,自引:0,他引:1
Farshid Ghassemi Mirko Vukcevic Le Xu Haiyan Zhou Gerhard Meissner Francesco Muntoni Heinz Jungbluth Francesco Zorzato Susan Treves 《Cell calcium》2009,45(2):192-197
Ryanodine receptors plays a crucial role in skeletal muscle excitation–contraction coupling by releasing calcium ions required for muscle contraction from the sarcoplasmic reticulum. At least three phenotypes associated with more than 100 RYR1 mutations have been identified; in order to elucidate possible pathophysiological mechanisms of RYR1 mutations linked to neuromuscular disorders, it is essential to define the mutation class by studying the functional properties of channels harbouring clinically relevant amino acid substitutions. In the present report we investigated the functional effects of the c.7304G > T RYR1 substitution (p.Arg2435Leu) found in a patient affected by central core disease. Both parents were heterozygous for the substitution while the proband was homozygous. We characterized Ca2+ homeostasis in myoD transduced myotubes from controls, the heterozygous parents and the homozygous proband expressing the endogenous mutation. We also expressed the recombinant mutant channels in heterologous cells and characterized their [3H]ryanodine binding and single channel properties. Our results show that the p.Arg2435Leu substitution affects neither the resting [Ca2+], nor the sensitivity of the ryanodine receptor to pharmacological activators, but rather reduces the release of Ca2+ from intracellular stores induced by pharmacological activators as well as by KCl via the voltage sensing dihydropyridine receptor. 相似文献
994.
Dan M. Berger Nancy Torres Minu Dutia Dennis Powell Greg Ciszewski Ariamala Gopalsamy Jeremy I. Levin Kyung-Hee Kim Weixin Xu James Wilhelm YongBo Hu Karen Collins Larry Feldberg Steven Kim Eileen Frommer Donald Wojciechowicz Robert Mallon 《Bioorganic & medicinal chemistry letters》2009,19(23):6519-6523
As part of our research effort to discover B-Raf kinase inhibitors, we prepared a series of C-3 substituted N-(3-(pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)-3-(trifluoromethyl)benzamides. X-ray crystallography studies revealed that one of the more potent inhibitors (10n) bound to B-Raf kinase without forming a hinge-binding hydrogen bond. With basic amine residues appended to C-3 aryl residues, cellular activity and solubility were enhanced over previously described compounds of this class. 相似文献
995.
目的:探讨胆汁性肝纤维化引起的门静脉高压症(PHT)大鼠肠系膜组织内皮素B受体表达(ETBR)的变化.方法:取体重250±10 g左右的清洁级SD雄性大鼠30只,根据体重随机分为假手术组、模型组,模型制作采用胆总管结扎的方法造成大鼠胆汁性肝纤维化.术后2周和4周分别测定各组的门静脉压力,应用免疫组化和免疫印迹的方法观察肠系膜组织ETBR的表达.结果:术后2周和4周模型组门静脉压力显著升高,分别为11.89±1.38 mmHg和16.34±1.63 mmHg.免疫组化显示假手术组肠系膜组织ETBR主要见于细动脉内皮细胞,而模型组大鼠ETBR的表达不仅见于细动脉内皮细胞,细动脉平滑肌细胞和微动脉表达也很显著.免疫印迹发现假手术组肠系膜组织ETBR含量很少,模型组大鼠ETBR表达明显增多.结论:正常大鼠肠系膜血管组织ETBR表达较少,随着肝组织损伤加剧和PHT形成,肠系膜组织ETBR表达明显增加,可能参与胆汁性肝硬化PHT形成过程. 相似文献
996.
Florence Quesada Calvo Marianne Fillet Dr. Dominique de Seny Marie‐Alice Meuwis Raphael Maree Céline Crahay Geneviève Paulissen Natacha Rocks Maud Gueders Louis Wehenkel Marie‐Paule Merville Renaud Louis Jean‐Michel Foidart Agnes Noël Didier Cataldo 《Proteomics》2009,9(8):2163-2170
Asthma is a complex inflammatory disease of airways. A network of reciprocal interactions between inflammatory cells, peptidic mediators, extracellular matrix components, and proteases is thought to be involved in the installation and maintenance of asthma‐related airway inflammation and remodeling. To date, new proteic mediators displaying significant activity in the pathophysiology of asthma are still to be unveiled. The main objective of this study was to uncover potential target proteins by using surface‐enhanced laser desorption/ionization‐time of flight‐mass spectrometry (SELDI‐TOF‐MS) on lung samples from mouse models of allergen‐induced airway inflammation and remodeling. In this model, we pointed out several protein or peptide peaks that were preferentially expressed in diseased mice as compared to controls. We report the identification of different five proteins: found inflammatory zone 1 or RELMα (FIZZ‐1), calcyclin (S100A6), clara cell secretory protein 10 (CC10), Ubiquitin, and Histone H4. 相似文献
997.
998.
999.
Shi-Xiao Xu Dong-Fang Cai Feng-Quan Tan Yan-Ni Fang Kai-Dong Xie Jude W. Grosser Wen-Wu Guo 《Plant Cell, Tissue and Organ Culture》2014,119(3):511-522
Citrus somatic hybrids produced in the past years provide a novel opportunity to study the immediate effects of allopolyploidization on genome structure and methylation. Here, we present a first attempt to investigate the alterations in genome structure and methylation in three sets of citrus somatic allotetraploids and their diploid parents using amplified fragment length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP) techniques. Our results indicate that all the allotetraploids mainly have the AFLP and MSAP banding patterns containing specific bands from both parents plus some alterations. The incidences of the AFLP polymorphic bands in allotetraploids show a range from 4.61 to 7.88 %, while from 12.50 to 15.67 % of the sites are methylated. In addition, the proportions of callus-parent-specific DNA structure and methylation alterations are much greater than those of leaf-parent-specific alterations in the somatic hybrids. Furthermore, we find that the somatic hybrids take on a greater divergence from the callus parent and a closer relationship to leaf parent in all groups of plants by dendrogram analysis based on AFLP or MSAP data. Taken together, our results suggest that somatic hybrids are very useful in elucidating the immediate changes that occur in newly synthesized allotetraploid. 相似文献
1000.
Longgen Xu Peng Han Yong Liu Hongwei Wang Yirong Yang Feng Qiu Wanling Peng Ligong Tang Jing Fu Xiaofeng Zhu Youhua Zhu 《Cell biochemistry and biophysics》2014,68(1):173-179
Even though the incidence of pregnancies in the female recipients is lower and also chronic renal disease in male patients is associated with impaired spermatogenesis, the health of the children born to these patients was not studied. In this report, we discuss information on the growth and development of offspring of 248 male and female kidney recipient patients. Physical and routine clinical measurements of the 252 offspring (129 male and 123 female) born to these transplantation patients were made along with the intelligence tests. In some of these children chest X-ray and immune indices were assessed. Among the recipients, 219 males fathered 223 children with an average birth weight of 3,255 ± 374 g and 29 female recipients gave birth to 29 children with an average birth weight of 2,923 ± 551. While most of these children were normal, we noticed a case of soft double toe, a case of short tongue tie, five cases of marginal mental retardation, three cases of proteinuria, six cases of microscopic hematuria, 15 cases of low hemoglobin, and 21 cases with recurrent respiratory tract infections. We conclude that kidney transplantation has no significant impact on the growth, development, health, and intelligence of the offspring born to recipients. 相似文献