title: Simple screening method for improving membrane protein thermostability.
abstract: Biochemical and biophysical analysis on integral membrane proteins often requires monodisperse and stable protein samples. Here we describe a method to characterize protein thermostability by measuring its melting temperature in detergent using analytical size-exclusion chromatography. This quantitative method can be used to screen for compounds and conditions that stabilize the protein. With this technique we were able to assess and improve the thermostability of several membrane proteins. These conditions were in turn used to assist purification, to identify protein ligand and to improve crystal quality.
authors.affiliation: The Helen L. and Martin S. Kimmel Center for Biology and Medicine at the Skirball Institute of Biomolecular Medicine, Department of Cell Biology, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
authors.forename: Romina
authors.initials: R
authors.lastname: Mancusso
authors.forename: Nathan K
authors.initials: NK
authors.lastname: Karpowich
authors.forename: Bryan K
authors.initials: BK
authors.lastname: Czyzewski
authors.forename: Da-Neng
authors.initials: DN
authors.lastname: Wang
mesh_terms: D027321:Anion Transport Proteins / Q000737:chemistry* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D005960:Glucosides / Q000737:chemistry; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification; D010711:Phosphatidate Phosphatase / Q000737:chemistry* / Q000302:isolation & purification; D055550:Protein Stability; D012995:Solubility; D044366:Transition Temperature
chemical_list: D027321:Anion Transport Proteins; D001426:Bacterial Proteins; D005960:Glucosides; D008565:Membrane Proteins; C040358:dodecyl maltoside; C112948:lipid phosphate phosphatase; D010711:Phosphatidate Phosphatase