Urea can inhibit efficient reduction and alkylation of protein dimers in solution demonstrated by the beta subunit of alpha glucosidase II

Meena Kumari; Antje Anji

doi:10.1016/j.ab.2018.10.008

Urea can inhibit efficient reduction and alkylation of protein dimers in solution demonstrated by the beta subunit of alpha glucosidase II

Anal Biochem. 2019 Feb 1:566:20-22. doi: 10.1016/j.ab.2018.10.008. Epub 2018 Oct 10.

Authors

Meena Kumari¹, Antje Anji²

Affiliations

¹ Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, 231Coles Hall, Manhattan, KS, 66506, USA. Electronic address: mkumari@vet.ksu.edu.
² Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, 231Coles Hall, Manhattan, KS, 66506, USA. Electronic address: aanji@vet.ksu.edu.

PMID: 30312619
DOI: 10.1016/j.ab.2018.10.008

Abstract

Protein reduction and alkylation is routinely used for analysis of protein dimers and protein complexes in cell fractions using two dimensional gel electrophoresis and mass spectrometry. To resolve the heterogeneity of a high molecular weight protein band that is highlighted by an antibody to the beta subunit of alpha glucosidase II (GIIβ), we performed reduction and alkylation of cytosolic proteins extracted from mouse brain. The presence of urea in the reduction/alkylation buffer inhibited the chemical processes. It is thus recommended that protein reduction/alkylation be performed both in the presence and absence of urea for the separation of mono-/hetero-mers.

Keywords: Protein dimer; Protein reduction; Urea; Western blotting.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alkylation
Animals
Brain Chemistry
Electrophoresis, Gel, Two-Dimensional / methods
Mass Spectrometry / methods
Mice
Protein Multimerization*
Urea / chemistry*
alpha-Glucosidases / chemistry*
alpha-Glucosidases / metabolism*

Substances

Urea
alpha-Glucosidases