相关实验视频
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Chromatographic Purification of Highly Active Yeast Ribosomes
Published on: October 24, 2011
通过纯化的酵母Sec复合体和没有膜的Kar2p进行蛋白质运输
K E Matlack1, K Plath, B Misselwitz
1Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
概括
酵母体内质网膜蛋白转位使用Sec复合体和Kar2蛋白. 该Sec复合物通过其道促进基质运动,由Kar2p和ATP水解提供动力.
科学领域:
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
- 蛋白质的运输方式
背景情况:
- 在细胞功能中,通过内细胞网膜 (ER) 进行翻译后蛋白质转移至关重要.
- 在酵母中,这种过程涉及七个组成部分的Sec复合体和光内Kar2蛋白 (Kar2p).
研究的目的:
- 阐明由Sec复合体和Kar2p介导的蛋白质转位的机制.
- 调查Kar2p和ATP水解在通过Sec复合道的基质运动中的作用.
主要方法:
- 使用了在洗剂溶液中复制的纯化Sec复合物.
- 对 Kar2p 和 ATP 的反应监测基质结合和转位.
- 在缺乏脂质双层的情况下研究过转位.
主要成果:
- 蛋白转位基质以信号序列依赖的方式与Sec复合体的细胞质面结合.
- Kar2p与光面相互作用,将ATP水解以驱动基质运动.
- 基质转位通过Sec复合体形成的通道发生,独立于脂质双层.
结论:
- 该Sec复合体形成了一个通道,促进蛋白质在ER膜上的转移.
- Kar2p作为一个电机,利用ATP水解来积极地通过Sec复杂道移动基质.
- 脂质双层对于Sec.复合体的转位通道功能来说并不重要.
相关概念视频
Overview of Secretory Vesicles
Secretory vesicles, also known as dense core vesicles (DCVs), are membrane-bound vesicles that transport secretory proteins, such as hormones or neurotransmitters. Regulated secretory vesicles transport proteins from the trans-Golgi network to the exterior of the cell. Proteins present in regulated secretory vesicles are required to be rapidly exocytosed in large amounts upon a specific stimulus.
Various proteins regulate the aggregation of molecules inside the secretory vesicles. Chromogranins...
Various proteins regulate the aggregation of molecules inside the secretory vesicles. Chromogranins...
Overview of Protein Sorting and Transport
Eukaryotic cells have different membrane-bound organelles with distinct protein requirements. The process by which proteins are targeted to a specific organelle is called protein sorting.
Protein sorting can be of two types: signal-based sorting and vesicle-based trafficking. In signal-based sorting, specific amino acid sequences called sorting signals target proteins to the proper location inside the cell either via gated transport or by protein translocation. In gated transport, folded...
Protein sorting can be of two types: signal-based sorting and vesicle-based trafficking. In signal-based sorting, specific amino acid sequences called sorting signals target proteins to the proper location inside the cell either via gated transport or by protein translocation. In gated transport, folded...
Nuclear Protein Sorting
Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Cotranslational Protein Translocation
Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...
Post-translational Translocation of Proteins to the RER
A sizable fraction of proteins destined for ER are first synthesized in the cell cytosol and then transported across the ER membrane–a process called post-translational translocation. Similar to cotranslationally translocated proteins, these proteins also use the Sec translocon complex to enter the ER lumen.
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
Protein Translocation Machinery on the ER Membrane
The translocon complex situated on the ER membrane is the main gateway for the protein secretory pathway. It facilitates the transport of nascent peptides into the ER lumen and their insertion into the ER membrane.
Sec61 protein conducting channel
In eukaryotes, the translocon complex comprises a core heterotrimeric translocator channel called the Sec61 complex. This channel includes three transmembrane proteins, Sec61α, Sec61β, and Sec61γ, and is the largest subunit of the translocon complex.
Sec61 protein conducting channel
In eukaryotes, the translocon complex comprises a core heterotrimeric translocator channel called the Sec61 complex. This channel includes three transmembrane proteins, Sec61α, Sec61β, and Sec61γ, and is the largest subunit of the translocon complex.

