本试验以蓖麻叶片为材料,克隆K+吸收蛋白基因Rc KUP 7,并对其进行生物信息学分析.结果显示,Rc KUP 7基因开放阅读框(Open Reading Frame,ORF)为2583 bp,编码860个氨基酸,蛋白分子量为95.149k D,p I值5.15.通过氨基酸组分分析得到含量最...本试验以蓖麻叶片为材料,克隆K+吸收蛋白基因Rc KUP 7,并对其进行生物信息学分析.结果显示,Rc KUP 7基因开放阅读框(Open Reading Frame,ORF)为2583 bp,编码860个氨基酸,蛋白分子量为95.149k D,p I值5.15.通过氨基酸组分分析得到含量最高的是亮氨酸(Leu),占该蛋白总摩尔质量的11.74%和分子质量的11.96%;含量最低的氨基酸为色氨酸(Trp),其摩尔质量与分子质量分别占总蛋白的1.16%和1.84%.进行多重序列比对和系统发育树分析可得出与麻枫树、木薯、橡胶树亲缘性较近.通过保守区蛋白分析可以得出该蛋白为钾离子转运蛋白家族的一员.通过亲水疏水性分析,可知该蛋白为疏水性蛋白.跨膜结构分析显示该蛋白具有10个蛋白跨膜结构,是典型的跨膜蛋白,并预测了该蛋白的三级结构.展开更多
Background： Disrupted Ca2＋ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis （UC）, but the underlying mechanisms are unknown. This study aimed to examine the alteration of col...Background： Disrupted Ca2＋ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis （UC）, but the underlying mechanisms are unknown. This study aimed to examine the alteration of colonic smooth muscle （SM） Ca2＋ signaling and Ca2＋ handling proteins in a rat model of dextran sulfate sodium （DSS）-induced UC. Methods： Male Sprague-Dawley rats were randomly divided into control （n = 18） and DSS （n = 17） groups. Acute colitis was induced by 5% DSS in the drinking water for 7 days. Contractility of colonic SM strips （controls, n = 8 and DSS, n = 7） was measured in an organ bath. Cytosolic resting Ca2＋ levels （n = 3 in each group） and Ca2＋ transients （n = 3 in each group） were measured in single colonic SM cells. Ca2＋ handling protein expression was determined by Western blotting （n = 4 in each group）. Differences between control and DSS groups were analyzed by a two-sample independent t-test. Results： Average tension and amplitude of spontaneous contractions of colonic muscle strips were significantly enhanced in DSS-treated rats compared with controls （1.25 ± 0.08 g vs. 0.96 - 0.05 g, P = 0.007; and 2.67 - 0.62 g vs. 0.52 ±0.10 g, P= 0.013）. Average tensions of carbachol-evoked contractions were much weaker in the DSS group （1.08 ±0.10 g vs. 1.80 ±0.19 g, P = 0.006）. Spontaneous Ca2＋ transients were observed in more SM cells from DSS-treated rats （15/30 cells） than from controls （5/36 cells）. Peak caffeine-induced intracellular Ca2＋ release was lower in SM cells of DSS-treated rats than controls （0.413 ±0.046 vs. 0.548 ±0.041, P = 0.033）. Finally, several Ca2＋ handling proteins in colonic SM were altered by DSS treatment, including sarcoplasmic reticulum calcium-transporting ATPase 2a downregulation and phospholamban and inositol 1,4,5-trisphosphate receptor 1 upregulation. Conclusions： Impaired intracellular Ca2＋ signaling of colonic SM, caused by alteration of Ca2＋ handing proteins, contribute to colonic dysmotility in DSS-induced UC.展开更多
文摘本试验以蓖麻叶片为材料,克隆K+吸收蛋白基因Rc KUP 7,并对其进行生物信息学分析.结果显示,Rc KUP 7基因开放阅读框(Open Reading Frame,ORF)为2583 bp,编码860个氨基酸,蛋白分子量为95.149k D,p I值5.15.通过氨基酸组分分析得到含量最高的是亮氨酸(Leu),占该蛋白总摩尔质量的11.74%和分子质量的11.96%;含量最低的氨基酸为色氨酸(Trp),其摩尔质量与分子质量分别占总蛋白的1.16%和1.84%.进行多重序列比对和系统发育树分析可得出与麻枫树、木薯、橡胶树亲缘性较近.通过保守区蛋白分析可以得出该蛋白为钾离子转运蛋白家族的一员.通过亲水疏水性分析,可知该蛋白为疏水性蛋白.跨膜结构分析显示该蛋白具有10个蛋白跨膜结构,是典型的跨膜蛋白,并预测了该蛋白的三级结构.
文摘Background： Disrupted Ca2＋ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis （UC）, but the underlying mechanisms are unknown. This study aimed to examine the alteration of colonic smooth muscle （SM） Ca2＋ signaling and Ca2＋ handling proteins in a rat model of dextran sulfate sodium （DSS）-induced UC. Methods： Male Sprague-Dawley rats were randomly divided into control （n = 18） and DSS （n = 17） groups. Acute colitis was induced by 5% DSS in the drinking water for 7 days. Contractility of colonic SM strips （controls, n = 8 and DSS, n = 7） was measured in an organ bath. Cytosolic resting Ca2＋ levels （n = 3 in each group） and Ca2＋ transients （n = 3 in each group） were measured in single colonic SM cells. Ca2＋ handling protein expression was determined by Western blotting （n = 4 in each group）. Differences between control and DSS groups were analyzed by a two-sample independent t-test. Results： Average tension and amplitude of spontaneous contractions of colonic muscle strips were significantly enhanced in DSS-treated rats compared with controls （1.25 ± 0.08 g vs. 0.96 - 0.05 g, P = 0.007; and 2.67 - 0.62 g vs. 0.52 ±0.10 g, P= 0.013）. Average tensions of carbachol-evoked contractions were much weaker in the DSS group （1.08 ±0.10 g vs. 1.80 ±0.19 g, P = 0.006）. Spontaneous Ca2＋ transients were observed in more SM cells from DSS-treated rats （15/30 cells） than from controls （5/36 cells）. Peak caffeine-induced intracellular Ca2＋ release was lower in SM cells of DSS-treated rats than controls （0.413 ±0.046 vs. 0.548 ±0.041, P = 0.033）. Finally, several Ca2＋ handling proteins in colonic SM were altered by DSS treatment, including sarcoplasmic reticulum calcium-transporting ATPase 2a downregulation and phospholamban and inositol 1,4,5-trisphosphate receptor 1 upregulation. Conclusions： Impaired intracellular Ca2＋ signaling of colonic SM, caused by alteration of Ca2＋ handing proteins, contribute to colonic dysmotility in DSS-induced UC.
