一种合的蛋白质和探针工程方法,用于选择性抑制和基于活动的探针标记caspases
Junpeng Xiao1, Petr Broz, Aaron W Puri
1Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA.
Journal of the American Chemical Society
|May 25, 2013
概括
研究人员为特定的标记和抑制设计了caspases. 这种新的方法允许精确研究在亡和炎症中的个体酶功能.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 囊酶是关键的囊蛋白酶,参与了亡和炎症.
- 由于保留了活跃地点和重叠的特点,研究个别的caspases具有挑战性.
研究的目的:
- 开发一种特定抑制和标记单个caspases的方法.
- 为了使caspase功能,激活和定位的详细调查.
主要方法:
- 通过将一个未保存的残留物转变为在活性部位附近的半氨酸来改造合成氨酸.
- 开发了一个带有电友的活点探测器,以准工程氨酸.
- 验证了使用caspase-8和caspase-1的方法.
主要成果:
- 工程化caspases在功能上与野生类型酶完全相同.
- 探测器专门抑制并标记了工程化卡斯帕斯.
- 证明了细胞内工程化caspase目标的直接成像.
结论:
- 这种方法允许特定的,不可逆转的抑制和标记工程 caspases.
- 这种方法有助于研究单个酶在细胞过程中扮演的角色,如细胞亡和炎症.
- 能够精确的成像caspase定位和活化在活细胞.
相关概念视频
03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
01:24Caspases
Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside cells.