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The Protein Journal

, Volume 35, Issue 5, pp 371–378 | Cite as

A Dynamic Interaction of Coomassie Dye with the Glycine Transporters N-termini

  • Anna Juhasova
  • Martina Baliova
  • Frantisek JurskyEmail author
Article

Abstract

Coomassie Brilliant Blue interacts with proteins and even though the interactions exhibit variation due to the amino acid content, reported dye interactions with individual proteins appear to be relatively stable. Here we report an atypical dynamic interaction of glycine transporters 1 and 2 N-termini with Coomassie dye, resulting in intramolecular interference with their Bradford assay. These proteins exhibit classic protein-Coomassie G-250 complex with absorption maximum at 595 nm, which within minutes starts to decrease and parallel increase of absorbance shoulders above 300 and 700 nm is observed. Interestingly, these effects are eliminated upon fusion of glycine transporters N-termini with glutathione S-transferase protein or by the presence of glutathione S-transferase or bovine serum albumin in the same solution. Circular dichroism data revealed largely unstructured character of glycine transporters N-termini, which suggests that dynamic properties of these protein- Coomassie complexes might be a signature of high flexibility and protein disorder. The assay might potentially reveal similar domains in other proteins and help to associate them with particular functions.

Keywords

Neurotransmitter transporters Glycine transporters Protein disorder Coomassie Brilliant Blue G-250 

Abbreviations

GlyT1

Glycine transporter 1

GlyT2

Glycine transporter 2

GST

Glutathione S-transferase

BSA

Bovine serum albumin

CBB

Coomassie Brilliant Blue

SDS

Sodium dodecyl sulfate

TEV

Tobacco etch virus

PCR

Polymerase chain reaction

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Notes

Acknowledgments

This work was supported by the Slovak grant agency VEGA, Grant 2/0086/13. The authors would like to thank Josef Houser PhD at the Biomolecular Interactions and Crystallization Core Facility of CEITEC for his expert technical assistance with obtaining CD spectra of proteins presented in this paper.

Compliance with Ethical Standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Anna Juhasova
    • 1
  • Martina Baliova
    • 1
  • Frantisek Jursky
    • 1
    Email author
  1. 1.Laboratory of Neurobiology, Institute of Molecular BiologySlovak Academy of SciencesBratislavaSlovakia

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