title: Peptergents: peptide detergents that improve stability and functionality of a membrane protein, glycerol-3-phosphate dehydrogenase.
abstract: Toward enhancing in vitro membrane protein studies, we have utilized small self-assembling peptides with detergent properties ("peptergents") to extract and stabilize the integral membrane flavoenzyme, glycerol-3-phosphate dehydrogenase (GlpD), and the soluble redox flavoenzyme, NADH peroxidase (Npx). GlpD is a six transmembrane spanning redox enzyme that catalyzes the oxidation of glycerol-3-phosphate to dihydroxyacetone phosphate. Although detergents such as n-octyl-beta-D-glucpyranoside can efficiently solubilize the enzyme, GlpD is inactivated within days once reconstituted into detergent micelles. In contrast, peptergents can efficiently extract and solubilize GlpD from native Escherichia coli membrane and maintain its enzymatic activity up to 10 times longer than in traditional detergents. Intriguingly, peptergents also extended the activity of a soluble flavoenzyme, Npx, when used as an additive. Npx is a flavoenzyme that catalyzes the two-electron reduction of hydrogen peroxide to water using a cysteine-sulfenic acid as a secondary redox center. The lability of the peroxidase results from oxidation of the sulfenic acid to the sulfinic or sulfonic acid forms. Oxidation of the sulfenic acid, the secondary redox center, results in inactivation, and this reaction proceeds in vitro even in the presence of reducing agents. Although the exact mechanism by which peptergents influence solution stability of Npx remains to be determined, the positive effects may be due to antioxidant properties of the peptides. Peptide-based detergents can be beneficial for many applications and may be particularly useful for structural and functional studies of membrane proteins due to their propensity to enhance the formation of ordered supramolecular assemblies.
authors.affiliation: Department of Molecular and Cell Biology, Brown University, Providence, Rhode Island 02912, USA. jiyeh@pitt.edu
authors.forename: Joanne I
authors.initials: JI
authors.lastname: Yeh
authors.forename: Shoucheng
authors.initials: S
authors.lastname: Du
authors.forename: Antoni
authors.initials: A
authors.lastname: Tortajada
authors.forename: Joao
authors.initials: J
authors.lastname: Paulo
authors.forename: Shuguang
authors.initials: S
authors.lastname: Zhang
mesh_terms: D002384:Catalysis; D003902:Detergents / Q000737:chemistry*; D016983:Enterococcus / Q000201:enzymology; D004795:Enzyme Stability; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology*; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D010544:Peroxidases / Q000737:chemistry / Q000378:metabolism; D012965:Sodium Chloride / Q000737:chemistry; D013997:Time Factors
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D010455:Peptides; D012965:Sodium Chloride; D005993:Glycerolphosphate Dehydrogenase; D010544:Peroxidases; C021845:NAD+ peroxidase