Literature DB >> 34541099

Thermostability Measurement of an α-Glucosidase Using a Classical Activity-based Assay and a Novel Thermofluor Method.

Karin Ernits1, Katrin Viigand1, Triinu Visnapuu1, Kristina Põšnograjeva1, Tiina Alamäe1.   

Abstract

α-glucosidases (including maltases and isomaltases) are enzymes which release glucose from a set of α-glucosidic substrates. Their catalytic activity, substrate specificity and thermostability can be assayed using this trait. Thermostability of proteins can also be determined using a high-throughput differential scanning fluorometry method, also named Thermofluor. We have shown that Thermofluor can also be applied to predict binding of substrates and inhibitors to a yeast α-glucosidase. The methods described here in detail were used in Viigand et al., 2016 .
Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC.

Entities:  

Keywords:  Differential scanning fluorometry; Glucose liquicolor; Isomaltase; Maltase; Maltase assay; Methylotrophic yeast; Ogataea polymorpha

Year:  2017        PMID: 34541099      PMCID: PMC8410288          DOI: 10.21769/BioProtoc.2349

Source DB:  PubMed          Journal:  Bio Protoc        ISSN: 2331-8325


  7 in total

1.  Thermodynamic analysis of ligand-induced changes in protein thermal unfolding applied to high-throughput determination of ligand affinities with extrinsic fluorescent dyes.

Authors:  Curtis J Layton; Homme W Hellinga
Journal:  Biochemistry       Date:  2010-12-03       Impact factor: 3.162

2.  Thermofluor-based high-throughput stability optimization of proteins for structural studies.

Authors:  Ulrika B Ericsson; B Martin Hallberg; George T Detitta; Niek Dekker; Pär Nordlund
Journal:  Anal Biochem       Date:  2006-08-10       Impact factor: 3.365

3.  The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability.

Authors:  Frank H Niesen; Helena Berglund; Masoud Vedadi
Journal:  Nat Protoc       Date:  2007       Impact factor: 13.491

Review 4.  Optimization of protein purification and characterization using Thermofluor screens.

Authors:  Stephane Boivin; Sandra Kozak; Rob Meijers
Journal:  Protein Expr Purif       Date:  2013-08-12       Impact factor: 1.650

5.  Genetic control of invertase formation in Saccharomyces cerevisiae. I. Isolation and characterization of mutants affecting sucrose utilization.

Authors:  R A Hackel
Journal:  Mol Gen Genet       Date:  1975-10-22

6.  Maltase protein of Ogataea (Hansenula) polymorpha is a counterpart to the resurrected ancestor protein ancMALS of yeast maltases and isomaltases.

Authors:  Katrin Viigand; Triinu Visnapuu; Karin Mardo; Anneli Aasamets; Tiina Alamäe
Journal:  Yeast       Date:  2016-04-21       Impact factor: 3.239

7.  The carbohydrate-active enzymes database (CAZy) in 2013.

Authors:  Vincent Lombard; Hemalatha Golaconda Ramulu; Elodie Drula; Pedro M Coutinho; Bernard Henrissat
Journal:  Nucleic Acids Res       Date:  2013-11-21       Impact factor: 16.971

  7 in total

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