A Ribosomal Misincorporation of Lys for Arg in Human Triosephosphate Isomerase Expressed in Escherichia coli Gives Rise to Two Protein Populations
2011

Ribosomal Error Yields Two Protein Populations

publication Evidence: moderate

Author Information

Author(s): Aguirre Beatriz, Costas Miguel, Cabrera Nallely, Mendoza-Hernández Guillermo, Helseth Donald L. Jr., Fernández Paulette, Tuena de Gómez-Puyou Marietta, Pérez-Montfort Ruy, Torres-Larios Alfredo, Gómez Puyou Armando

Primary Institution: Universidad Nacional Autónoma de México

Hypothesis

Can ribosomal misincorporation of lysine for arginine in human triosephosphate isomerase lead to the formation of two distinct protein populations?

Conclusion

The study demonstrates that ribosomal misincorporation can produce two proteins with different biochemical properties from the same gene sequence.

Supporting Evidence

  • The two proteins were present in nearly equimolar amounts under certain growth conditions.
  • Both proteins correspond to human triosephosphate isomerase as shown by mass spectrometry.
  • The mass difference of 28 Da is due to the substitution of a lysine for an arginine residue.
  • Expression in a strain with extra tRNA copies led to the exclusive production of the more stable protein.
  • Both proteins exhibited different thermal stability and susceptibility to denaturation.

Takeaway

Sometimes, when cells make proteins, they can make mistakes and end up with two slightly different versions of the same protein, which can behave differently.

Methodology

The study involved expressing human triosephosphate isomerase in E. coli, isolating the resulting proteins, and analyzing their properties using techniques like size exclusion chromatography and mass spectrometry.

Limitations

The study primarily focuses on a specific protein and may not generalize to all proteins expressed in E. coli.

Digital Object Identifier (DOI)

10.1371/journal.pone.0021035

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