title: Large-scale identification of membrane proteins with properties favorable for crystallization.
abstract: Membrane protein crystallography is notoriously difficult due to challenges in protein expression and issues of degradation and structural stability. We have developed a novel method for large-scale screening of native sources for integral membrane proteins that have intrinsic biochemical properties favorable for crystallization. Highly expressed membrane proteins that are thermally stable and nonaggregating in detergent solutions were identified by mass spectrometry from Escherichia coli, Saccharomyces cerevisiae, and Sus scrofa cerebrum. Many of the membrane proteins identified had been crystallized previously, supporting the promise of the approach. Most identified proteins have known functions and include high-value targets such as transporters and ATPases. To validate the method, we recombinantly expressed and purified the yeast protein, Yop1, which is responsible for endoplasmic reticulum curvature. We demonstrate that Yop1 can be purified with the detergent dodecylmaltoside without aggregating.
authors.affiliation: Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Jared
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Allison
authors.initials: A
authors.lastname: Kagawa
authors.affiliation: Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Kellie
authors.initials: K
authors.lastname: Kurasaki
authors.affiliation: Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Niloufar
authors.initials: N
authors.lastname: Ataie
authors.affiliation: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Il Kyu
authors.initials: IK
authors.lastname: Cho
authors.affiliation: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Qing X
authors.initials: QX
authors.lastname: Li
authors.affiliation: Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.|University of Hawaii Cancer Center, University of Hawaii at Manoa, Honolulu, Hawaii, 96822.
authors.forename: Ho Leung
authors.initials: HL
authors.lastname: Ng
mesh_terms: D000818:Animals; D003460:Crystallization / Q000379:methods*; D029968:Escherichia coli Proteins; D013058:Mass Spectrometry / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D013552:Swine
chemical_list: D029968:Escherichia coli Proteins; D008565:Membrane Proteins
keywords: crystallization; mass spectrometry; membrane proteins; proteomics