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BA Joughin C Liu DA Lauffenburger CW Hogue MB Yaffe 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2012,367(1602):2574-2583
Characterization of in vitro substrates of protein kinases by peptide library screening provides a wealth of information on the substrate specificity of kinases for amino acids at particular positions relative to the site of phosphorylation, but provides no information concerning interdependence among positions. High-throughput techniques have recently made it feasible to identify large numbers of in vivo kinase substrates. We used data from experiments on the kinases ATM/ATR and CDK1, and curated CK2 substrates to evaluate the prevalence of interactions between substrate positions within a motif and the utility of these interactions in predicting kinase substrates. Among these data, evidence of interpositional sequence dependencies is strikingly rare, and what dependency exists does little to aid in the prediction of novel kinase substrates. Significant increases in the ability of models to predict kinase-substrate specificity beyond position-independent models must come largely from inclusion of elements of biological and cellular context, rather than further analysis of substrate sequences alone. Our results suggest that, evolutionarily, kinase substrate fitness exists in a smooth energetic landscape. Taken with results from others indicating that phosphopeptide-binding domains do exhibit interpositional dependence, our data suggest that incorporation of new substrate molecules into phospho-signalling networks may be rate-limited by the evolution of suitability for binding by phosphopeptide-binding domains. 相似文献
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桃一点叶蝉及草间小黑蛛空间格局的地学统计学研究 总被引:6,自引:11,他引:6
应用地学统计学的原理和方法方法研究了不同时期桃-点叶蝉及草间小黑蛛种群的空间结构和空间相关性。结果表明,不同时期桃-点叶蝉种群的半变异函数曲线皆为球型,其空间格局为聚集型,变程在25.71-37.14m之间,草间小黑蛛种群的拟合并变异函数也表现为球型,呈聚集空间格局,空间变程在25.12-44.06m之间,并与桃-点叶蝉种群在数量和空间上有较强的追随关系,说明草间小黑蛛种群是桃一点叶蝉种群的优势种天敌。 相似文献
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DDTs(dichlorodiphenyltrichloroethane,1,1,1-三氯-2,2-双氯苯基乙烷)是一种典型的持久性有机污染物,曾在疟疾防治和农业除虫方面被广泛应用。虽然包括我国在内的很多国家已经禁止使用DDTs,但目前对环境中DDTs的检测发现它仍然广泛存在且具有新的输入源。DDTs的持续存在对近海生态系统和人类健康具有一定危害,因此它所造成的环境污染问题仍然值得关注。由于Rieske型芳香羟化双加氧酶能够起始多种持久性污染物的降解,过去的几十年里一直是芳香化合物降解领域的焦点。[目的] 为探讨联苯双加氧酶对DDTs的降解特性及机制,本研究选取了食异生素伯克霍尔德氏菌LB400(Burkholderia xenovorans)联苯双加氧酶及突变体对p,p''-DDT和o,p''-DDT的降解过程进行研究。[方法] 以BphAELB400为亲本,通过两步定点突变将283位的丝氨酸突变为蛋氨酸,获得突变体BphAES283M。通过比较亲本酶与突变体对DDTs的催化性能,模拟突变蛋白结构和分子对接等方法,探究其降解特性及机制。[结果] BphAELB400和突变体BphAES283M都无法降解对位的p,p''-DDT,但突变体BphAES283M可以代谢o,p''-DDT并产生2个立体异构体。对接p,p''-DDT的BphAELB400和BphAES283M的结构分析表明,BphAELB400和BphAES283M中p,p''-DDT的反应环均不与原晶体结构中的联苯反应环重合。而对接o,p''-DDT的BphAES283M的结构分析表明o,p''-DDT的反应环与晶体结构中的联苯反应环距离很近,且2、3位的碳原子与单核铁原子催化中心的距离在0.5 nm以内,此外,BphAES283M的催化腔表面积和体积比BphAELB400更大,这很可能有助于BphAES283M与o,p''-DDT的结合。[结论] 283位氨基酸是影响BphAELB400对DDTs的催化代谢能力的关键氨基酸残基,它可以通过调节反应碳原子与催化中心的距离以及催化腔的大小来影响底物特异性。本次研究进一步阐明了283位氨基酸残基的影响机理,为更有效修复DDTs污染提供理论依据和技术支持。 相似文献
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Pengju Zhang Jun Wang Hongyan Lang Weixia Wang Xiaohui Liu Haiyan Liu Chengcheng Tan Xintao Li Yumin Zhao Xinghong Wu 《Journal of cellular biochemistry》2019,120(5):8466-8474
MicroRNA-205 (miR-205) is involved in various physiological and pathological processes, but its biological function in follicular atresia remains unclear. In this study, we investigated miR-205 expression in mouse granulosa cells (mGCs) and analyzed its functions in primary mGCs by performing a series of in vitro experiments. Quantitative real-time polymerase chain reaction showed that miR-205 expression was significantly higher in early atretic follicles and progressively atretic follicles than in healthy follicles. miR-205 overexpression in mGCs significantly promoted apoptosis and caspase-3/9 activities, as well as inhibited estrogen (E2) release and cytochrome P450 family 19 subfamily A polypeptide 1 (CYP19A1, a key gene in E2 production) expression. Bioinformatics and luciferase reporter assays revealed that the gene encoding cyclic AMP response element (CRE)-binding protein 1 (CREB1) was a direct target of miR-205 in mGCs. CREB1 upregulation partially rescued the effects of miR-205 on apoptosis, caspase-3/9 activities, E2 production, and CYP19A1 expression on mGCs. These results indicate that miR-205 might play an important role in ovarian follicular development and provide new insights into follicular atresia 相似文献
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