课题基金 / 基金详情

AMINO ACID LABELING FOR LARGER PROTEINS METHYL GROUP LABELING

AMINO ACID LABELING FOR LARGER PROTEINS METHYL GROUP LABELING
较大蛋白质的氨基酸标记 甲基组标记
批准号:
6120852
负责人:
RODOLFO MARTINEZ
金额:
$7.79万
依托单位国家:
美国
项目类别:
财政年份:
1999
资助国家:
美国
项目状态:
已结题
起止时间:
1999-01-15 至 2000-01-14

项目摘要

项目成果

RODOLFO MARTINEZ的其他基金

相似基金

相关文献

中文摘要
翻译
SIR提供3,4,6二氯- 5,15纳米嘧啶;20毫克
英文摘要
The SIR provided 3,4,6 dichloro-5 15 Namino-pyrimidine; 20mg 6,2'deoxy[9J5N]adenosine; 20mg I a tellurocystine; 20mg 93-94% "Selenium; 87mg Selenomethionine containing proteins analyzed by multiwavelength anomalous diffraction (MAD) provide a facile means of addressing the phase problem, whose solution is necessary to determine protein structures by X-ray crystallography [Hendrickson, W.A. (1991) Science, 254, 51-58]. Since this method requires synchrotron radiation, we sought to incorporate a true heavy atom into protein, allowing the solution of the phase problem by more traditional methods of data collection. Media containing Te-Met alone or Te-Met with low levels of Met failed to sustain growth of a methionine auxotroph of Escherichia coli carrying the dihydrofolate reductase expression vector. Growth of the organism to stationary phase and incorporation of Te-Met was observed when the culture was initiated in media containing minimal Met levels and Te-Met was added after induction with IPTG. The purified enzyme exhibited properties similar to those of the native enzyme. Atomic absorption spectroscopy and amino acid analysis indicated that 40% of the methionines were replaced with Te-Met. Sequence analysis did not indicate significant levels of replacement in the first three sites (1, 16, and 20), suggesting that Te-Met was present only in the last two sites (42 and 92).Crystals of this enzyme were grown in the presence of methotrexate and were isomorphous with crystals of wild-type DHFR. Difference Fourier maps and restrained least-squares refinement showed no substitution at the first three methionines, while incorporation was seen at positions 42 and 92.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
SYNTHESIS OF [1,3_2 H4, 2 13C] GLYCEROL, & [1,2,3_2 H31 1,3 13 C21GLYCEROL
AMINO ACID LABELING FOR LARGER PROTEINS METHYL GROUP LABELING
SYNTHESIS OF LABELED PRECURSORS
SYNTHESIS OF LABELED PRECURSORS
海外基金