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Related Concept Videos

ABC Transporters: Importer01:27

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ATP-binding cassette or ABC transporters are a class of ATP-driven pumps that hydrolyze ATP to move solutes across the membrane. They can be grouped into importers and exporters. While exporters are present in all domains of life, importers exist only in bacteria and some plants.
In bacteria, based on the number of transmembrane helices and the chemical nature of their substrates, the ABC importers can be divided into three types:
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ABC Transporters: Exporter01:31

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ATP-binding cassette or ABC transporter is the largest superfamily of integral membrane proteins. The transporters have transmembrane-binding domains (TMDs) and nucleotide-binding domains (NBDs). The TMDs are specific to their substrates, whereas the NBDs are similar to engines that complete ATP hydrolysis to complete the substrate transport. They can be full transporters consisting of two TMDs and NBDs, half transporters with one TMD and NBD, while some encoded with a single TMD or NBD are...
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Updated: Jun 28, 2025

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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PySupercharge: a python algorithm for enabling ABC transporter bacterial secretion of all proteins through amino acid

Yerin Kim1, Danny Kim1, Nguyen-Mihn Hieu1

  • 1Department of Chemistry and Biology, Korea Science Academy of Korea Advanced Institute of Science and Technology, Busan, South Korea.

Microbial Cell Factories
|April 20, 2024
PubMed
Summary

Protein secretion via ATP-binding cassette (ABC) transporters is hindered by excessive positive charge. We developed PySupercharge to modify protein charge, enabling secretion of previously non-secretable proteins.

Keywords:
ABC TransporterProtein productionPython AlgorithmSecretionSupercharging

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Background:

  • Bacterial protein production and secretion via ATP-binding cassette (ABC) transporters offer high capacity and efficiency.
  • Certain proteins fail to secrete due to excessive positive charge in specific amino acid sequence regions.
  • Removing excessive positive charge is key to enabling universal protein secretion through ABC transporters.

Purpose of the Study:

  • To establish 'linear charge density' as a criterion for protein secretion possibility through ABC transporters.
  • To develop a computational tool for enabling the secretion of proteins with high linear charge density.
  • To demonstrate the applicability of the developed method to various non-secretable proteins.

Main Methods:

  • Introduced 'linear charge density' as the primary criterion for assessing protein secretion feasibility.
  • Developed a novel algorithm, PySupercharge, for analyzing and modifying protein sequences.
  • PySupercharge selectively converts positively charged amino acids to neutral or negatively charged ones based on linear charge density analysis.

Main Results:

  • Confirmed that linear charge density accurately predicts protein secretion through ABC transporters.
  • Successfully applied the criterion to non-secretable proteins including SARS-CoV-2 spike proteins, botulinum toxin light chain, and human growth factors.
  • PySupercharge demonstrated efficacy in enabling the secretion of these challenging proteins.

Conclusions:

  • PySupercharge minimizes functional and structural stability loss by utilizing sequence conservation data.
  • The PySupercharge tool is available via a web server for research applications.
  • Validated PySupercharge's effectiveness in secreting previously non-secretable proteins, recommending it for future protein production research.