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L-pyrrolysine Chemical and Structural Biology

L-吡咯赖氨酸化学和结构生物学

基本信息

批准号：
7270620
负责人：
Michael K Chan
金额：
$ 27.64万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-05 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7270620
关键词：
Address Amber Amines Amino Acids Amino Acyl-tRNA Synthetases Binding Biochemical Biochemistry Biotin Chemicals Codon Nucleotides Complex Corrinoids Coumarins Dimethylamines Environment Fluorescence Fluorescent Probes Funding Future Goals Grant Hydroxylamine Imines In Vitro Laboratories Lead Ligation Lysine Maps Mediating Methanosarcina Methylamines Methyltransferase Modification Names Nature Nitrogen Object Attachment Process Property Proteins Radioactive Range Recombinant Proteins Recombinants Role Series Site-Directed Mutagenesis Specificity Structure Substrate Specificity Techniques Technology Transfer RNA United States National Institutes of Health alpha benzopyrone analog base chemical synthesis cofactor delta-1-pyrroline-5-carboxylate design dimethylamine electron density functional group genetic evolution in vivo insight intracellular protein transport methylamine monomethylamine methyltransferase novel protein expression protein localization location protein purification pyrrolysine structural biology tool

项目摘要

DESCRIPTION (provided by applicant): During the previous funding cycle, we determined that a UAG-codon, located in the M. barkeri monomethylamine methyltransferase, encoded a new amino acid. Based on its electron density in the native and hydroxylamine complexes, we determined that the amino acid is 4-methyl-(4R,5R)-pyrroline-5- carboxylate, attached to the epsilon nitrogen of lysine. Based on its composition, we named this amino acid, L-pyrrolysine. The subsequent identification, by the Krzycki group, of an amber decoding tRNA(Pyl), and a novel L-pyrrolysyl-tRNA synthetase PylS, that loads L-pyrrolysine onto tRNA(Pyl) make a strong case for Lpyrrolysine being the 22nd genetically-encoded amino acid in nature. During this funding cycle, we propose to continue our efforts in understanding L-pyrrolysine's functional role and its mechanism of incorporation into proteins. This will include: (1) structural studies of L-pyrrolysine containing proteins, (2) structural studies involved in L-pyrrolysine incorporation into protein, and (3) chemical synthesis of L-pyrrolysine and its analogs.

描述（由申请人提供）：在上一个资助周期，我们确定了一个UAG密码子，位于M。巴克氏单甲胺甲基转移酶，编码一个新的氨基酸。基于其在天然和羟胺络合物中的电子密度，我们确定氨基酸是4-甲基-（4 R，5 R）-吡咯啉-5-羧酸酯，连接到赖氨酸的N-端氮。基于其组成，我们将这种氨基酸命名为L-吡咯赖氨酸。Krzycki小组随后鉴定了琥珀解码tRNA（Pyl）和将L-吡咯赖氨酸装载到tRNA（Pyl）上的新型L-吡咯赖氨酰-tRNA合成酶PylS，这为L-吡咯赖氨酸是自然界中第22种遗传编码的氨基酸提供了有力的证据。在本资助周期内，我们建议继续努力了解L-吡咯赖氨酸的功能作用及其掺入蛋白质的机制。这将包括：（1）含L-吡咯赖氨酸的蛋白质的结构研究，（2）涉及L-吡咯赖氨酸掺入蛋白质的结构研究，和（3）L-吡咯赖氨酸及其类似物的化学合成。