title: Analyses of in vitro nonenzymatic glycation of normal and variant hemoglobins by MALDI-TOF mass spectrometry.
abstract: MALDI-TOF mass spectrometry is used here to differentiate different glycoisoforms of normal and variant hemoglobins (Hbs) in nonenzymatic in vitro glycation. Single, double, and/or multiple glycation of the α-globin, β-globin, and/or γ-globin is observed. Different glycation rates are observed for various Hbs, and the normal Hb A has the slowest rate. Although the Hb A is relatively stable upon condensation with glucose at 37°C, the variants Hb C, Hb E, Hb F, Hb Leiden, and Hb San Diego are less stable. In addition, data reveal that the number of glucose attached/Hb molecule (state of glycation) increases with longer incubation time, higher glucose concentration, and higher temperature. The pH dependence of the state of glycation is more complex and varies for different Hbs. Although pH has little effect on the state of glycation for Hb C, Hb E, and Hb Leiden, it increases for Hb A and Hb F upon changing the pH of the solution from phosphate buffer saline (pH 7.4) to carbonate buffer (pH 10). Results obtained in this study could lead to the inference that the linkage of Hbs with glucose occurs in diabetic conditions in vivo (37°C, ∼neutral pH, ∼0.007 M glucose), and the state of glycation is more severe in the individuals who carry abnormal Hbs.
authors.affiliation: Protein Research Laboratory, Research Resources Center, University ofIllinois at Chicago, Chicago, Illinois 60612, USA. boblee@uic.edu
authors.forename: Bao-Shiang
authors.initials: BS
authors.lastname: Lee
authors.forename: G D Lasanthi P
authors.initials: GD
authors.lastname: Jayathilaka
authors.forename: Jin-Sheng
authors.initials: JS
authors.lastname: Huang
authors.forename: Loyda N
authors.initials: LN
authors.lastname: Vida
authors.forename: George R
authors.initials: GR
authors.lastname: Honig
authors.forename: Shalini
authors.initials: S
authors.lastname: Gupta
mesh_terms: D003920:Diabetes Mellitus / Q000097:blood; D005947:Glucose / Q000737:chemistry; D006031:Glycosylation; D006454:Hemoglobins / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006454:Hemoglobins; D005947:Glucose