相关实验视频

Updated: Jan 12, 2026

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Published on: March 9, 2010

AP-4辅助蛋白质tepsin表现出对LC3B的多价值结合

Cameron I Cohen¹, Amy K Kendall¹, Natalie S Wallace¹

¹Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.

Advances in biological regulation

|November 6, 2025

概括

素是一种适应蛋白4 (AP-4) 囊泡蛋白,通过四种基因与LC3B结合. 这种多价值性使得特素能够根据LC3B度动态调节结合强度,影响ATG9A贩运和自细胞成熟.

科学领域:

细胞生物学细胞生物学
分子生物学分子生物学
生物化学生物化学

背景情况:

素是适应蛋白4 (AP-4) 涂层囊泡中的辅助蛋白,对于跨戈尔吉网络 (TGN) 的货物贩运至关重要.
AP-4囊泡处理货物,如ATG9A,这是一个脂质混杂酶,对自细胞成熟至关重要.
素已知的LC3相互作用区域 (LIR) 动机影响ATG9A分布和自细胞形态.

研究的目的:

为了研究tepsin和LC3B.之间的分子相互作用.
描述tepsin-LC3B相互作用的结合动力学和热力学.
了解特普辛的多价值性如何促进其在囊泡贩运和自中的功能.

主要方法:

使用AlphaFold Multimer进行计算建模.
生物化学和生物物理实验,包括生物层干涉测量 (BLI).
测量tepsin-LC3B结合的热力学和动力学特性.

主要成果:

除了正规的LIR外,在tepsin的无序区域中还发现了三个额外的LC3B结合动机.
所有四个图案都独立地与LC3B LIR对接点 (LDS) 结合.
为了在体外取消LC3B结合,必须对所有四种基因进行突变;特普辛可能同时结合两个LC3B分子.

结论:

特普辛利用通过四种不同的LC3B结合基因的多价值性来调节结合强度.
这种多价值性使得特普辛能够对细胞LC3B度有动态反应.
这些发现为AP-4囊泡贩运和自细胞成熟的调节提供了洞察力.

更多相关视频

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Published on: October 3, 2019

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells

Published on: January 26, 2017

相关概念视频

Tail-anchoring of Proteins in the ER Membrane

Tail-anchoring of Proteins in the ER Membrane

Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...

Protein Complexes with Interchangeable Parts

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...

Protein Translocation Machinery on the ER Membrane

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...