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Related Concept Videos

Insertion of Multi-pass Transmembrane Proteins in the RER01:29

Insertion of Multi-pass Transmembrane Proteins in the RER

The rough ER membrane synthesizes, assembles, and embeds transmembrane proteins in diverse topologies. These proteins function as transporters or channels and can remain in the ER membrane or are sent to the Golgi complex, lysosome, and cell membrane.
The multipass transmembrane proteins are the type IV integral membrane proteins with multiple topogenic sequences determining their spatial arrangement in the ER membrane. Nearly all multipass proteins lack a cleavable signal sequence and use...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
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Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Cotranslational Protein Translocation01:20

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Sec61 channel partners for cotranslational translocation
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Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
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Related Experiment Video

Updated: May 22, 2026

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
10:56

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

Published on: May 17, 2014

Translation levels control multi-spanning membrane protein expression.

Hok Seon Kim1, James A Ernst, Cecilia Brown

  • 1Department of Antibody Engineering, Genentech Inc., South San Francisco, California, United States of America.

Plos One
|May 8, 2012
PubMed
Summary
This summary is machine-generated.

Controlling translation initiation rates is key for high-level expression of membrane proteins. Moderate rates improve protein synthesis, targeting, and accumulation in E. coli.

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Published on: May 17, 2014

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Eukaryotic multi-spanning membrane proteins are challenging to express at high levels.
  • Understanding limitations in membrane protein expression is crucial for research.

Purpose of the Study:

  • To investigate the impact of translation levels on the expression of human membrane proteins in E. coli.
  • To identify rate-limiting steps in membrane protein synthesis and accumulation.

Main Methods:

  • Analysis of translation initiation rates for four human membrane proteins (CD20, RA1c, EG-VEGFR1, Patched 1) in E. coli.
  • Evaluation of protein synthesis, targeting, insertion, and folding efficiency.

Main Results:

  • Excessive translation initiation rates block protein synthesis and prevent high-level accumulation.
  • Moderate translation rates facilitate coupled peptide synthesis and membrane targeting, enhancing expression.
  • Translation initiation rates critically influence the targeting, insertion, and folding of integral membrane proteins.

Conclusions:

  • Translation initiation rate is a critical determinant for successful high-level expression of eukaryotic membrane proteins in E. coli.
  • Optimizing translation rates can overcome expression limitations and improve yields of membrane proteins for study.