照片诱导的,特定于家族的,选择性地点切割TIM-桶蛋白的TIM-桶蛋白
Nicola Floyd1, Neil J Oldham, Christopher J Eyles
1Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford, OX1 3TA, UK.
Journal of the American Chemical Society
|September 3, 2009
概括
研究人员发现了一种使用光线切割蛋白质的新方法. 这种光化学蛋白质分裂方法在特定的His-Trp动机上选择性地分解蛋白质,从而使蛋白质组学和蛋白质净化中的新应用成为可能.
科学领域:
- 生物化学 生物化学
- 摄影化学的使用.
- 分子生物学分子生物学
背景情况:
- 控制的蛋白质分裂对于翻译后修改,蛋白质组学和蛋白质工程至关重要.
- 现有的非酶,特定位点的蛋白质分裂方法是有限的.
- GH1家族TIM-桶蛋白是一种具有结构操纵潜力的酶类.
研究的目的:
- 开发一种新的光化学方法,用于特定位置的蛋白质裂变.
- 为了研究一个His-Trp (HW) 图案的裂变机制.
- 探索这个图案在蛋白质净化和工程中的实用性.
主要方法:
- 用紫外线 (240-308 nm) 照射GH1家族TIM桶蛋白中的His-Trp (HW) 基因.
- 使用质谱和埃德曼降解分析蛋白质碎片的分析.
- 将可光切割的图案纳入聚变蛋白中,用于亲和力净化实验.
主要成果:
- 在暴露于光线时,实现了His-Trp图案的光化学,选择性切割.
- 裂变清洁释放的N端初级胺和C端胺片段.
- 可光切割的图案成功转移到融合蛋白中,使光响应亲和力净化成为可能.
结论:
- 已经建立了一个新的,非酶的,光化学方法,用于特定位点的蛋白质裂变.
- 这种方法提供了一种清洁和可控的方式来操纵蛋白质结构.
- 可光切割的His-Trp图案在蛋白质组学,蛋白质测序和光响应亲和力净化方面具有潜在的应用.
更多相关视频
11:33Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
Published on: December 17, 2013
08:13Spatiotemporal Subcellular Manipulation of the Microtubule Cytoskeleton in the Living Preimplantation Mouse Embryo using Photostatins
Published on: November 30, 2021
相关概念视频
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
Maxam-Gilbert Sequencing
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
Translocation of Proteins into the Mitochondria
Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
Translesion DNA Polymerases
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
