Urea can inhibit efficient reduction and alkylation of protein dimers in solution demonstrated by the beta subunit of alpha glucosidase II.
Identifieur interne : 000334 ( Main/Exploration ); précédent : 000333; suivant : 000335Urea can inhibit efficient reduction and alkylation of protein dimers in solution demonstrated by the beta subunit of alpha glucosidase II.
Auteurs : Meena Kumari [États-Unis] ; Antje Anji [États-Unis]Source :
- Analytical biochemistry [ 1096-0309 ] ; 2019.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Urea, alpha-Glucosidases.
- chemical , metabolism : alpha-Glucosidases.
- methods : Electrophoresis, Gel, Two-Dimensional, Mass Spectrometry.
- Alkylation, Animals, Brain Chemistry, Mice, Protein Multimerization.
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.
DOI: 10.1016/j.ab.2018.10.008
PubMed: 30312619
Affiliations:
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Le document en format XML
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<term>Protein Multimerization</term>
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<term>alpha-Glucosidases (chemistry)</term>
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<term>Biochimie de l'encéphale</term>
<term>Multimérisation de protéines</term>
<term>Souris</term>
<term>Spectrométrie de masse ()</term>
<term>Urée ()</term>
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<term>alpha-Glucosidase (métabolisme)</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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