title: Surface and interface beta-chain residues synergistically affect hemoglobin assembly.
abstract: Homo- and heterotetramer formations of beta112 variants (beta(112Cys-->Asp), beta(112Cys-->Ser), beta(112Cys-->Thr), and beta(112Cys-->Val)) of hemoglobin were characterized in the presence and absence of beta(16Gly-->Asp) in vitro. In all cases an alteration in overall surface charge (beta(16Gly-->Asp)) decreased the beta(4) homotetramer stability (association constants as determined by gel-permeation chromatography) albeit to differing extents. In contrast, competition experiments of hemoglobin subunits showed that heterotetramer formation was promoted by this substitution. Order of increase in tetramer formation by the additional negative surface charge in the beta112 variants was as follows: Hb betaG16D, C112D > Hb betaG16D, C112S > Hb betaG16D > Hb G16D, C112T > Hb betaG16D, C112V. Thus, the overall surface charge of the beta chain and its contribution to electrostatic interaction in these instances appear to act in synergy with alpha(1)beta(1) interface residues to affect the assembly of hemoglobin molecules.
authors.affiliation: The Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104, USA.
authors.forename: T
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: M J
authors.initials: MJ
authors.lastname: McDonald
authors.forename: K
authors.initials: K
authors.lastname: Adachi
mesh_terms: D019943:Amino Acid Substitution; D014644:Genetic Variation; D005914:Globins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D016297:Mutagenesis, Site-Directed; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D055672:Static Electricity
chemical_list: D006454:Hemoglobins; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D005914:Globins