title: Conformational stability of adsorbed insulin studied with mass spectrometry and hydrogen exchange.
abstract: A new method is described for direct monitoring of the conformational stability of proteins that are physically adsorbed from solution onto a solid substrate. The adsorption-induced conformational changes of insulin are studied using a combination of hydrogen/deuterium (H/D) exchange and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The effect of the surface hydrophobicity on the adsorption-induced conformational changes in the insulin structure is probed by adsorbing insulin on a hydrophilic silica and a hydrophobic methylated silica surface before subjecting the insulin molecules to the isotopic exchange process. The present study describes the experimental procedure of this new application of MALDI. Results show that insulin is more highly and more irreversibly adsorbed to a hydrophobic methylated silica surface than to a hydrophilic silica surface. Hydrogen-exchange experiments clearly demonstrate that the strong interaction of insulin with the hydrophobic surface is accompanied by a strong increase in the H/D-exchange rates and thus in a reduction in the insulin native structural stability. In contrast, H/D-exchange rates of insulin are somewhat reduced upon adsorption on silica from solution.
authors.affiliation: Division of Ion Physics, Angström Laboratory, Uppsala University, Sweden. Jos.Buijs@angstrom.uu.se
authors.forename: J
authors.initials: J
authors.lastname: Buijs
authors.forename: C
authors.initials: C
authors.lastname: Costa Vera
authors.forename: E
authors.initials: E
authors.lastname: Ayala
authors.forename: E
authors.initials: E
authors.lastname: Steensma
authors.forename: P
authors.initials: P
authors.lastname: Håkansson
authors.forename: S
authors.initials: S
authors.lastname: Oscarsson
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D003903:Deuterium / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D007328:Insulin / Q000737:chemistry*; D007700:Kinetics; D011487:Protein Conformation; D012822:Silicon Dioxide; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013499:Surface Properties
chemical_list: D007328:Insulin; D012996:Solutions; D012822:Silicon Dioxide; D006859:Hydrogen; D003903:Deuterium