Effects of pH, 2,3-diphosphoglycerate and salts on gelation of sickle cell deoxyhemoglobin

Robin W. Briehl, Sandra Ewert

Research output: Contribution to journalArticle

82 Scopus citations

Abstract

Gelation of fully deoxygenated sickle cell hemoglobin was assayed by (1) determination of the temperature at which viscosity increased sharply and (2) a high-speed sedimentation equilibrium method in which three zones are seen. These are a pre-gelation zone, a narrow transition zone exhibiting aggregation, followed by a phase change and a zone of gelation. Only the first zone is seen with deoxyhemoglobin A and CO hemoglobins A and S up to about 0·35 g protein/ml. Minimal gelling temperatures by the viscosity method and, by ultracentrifugation, minimal gelling concentrations determined at the onset of aggregation and at the phase change showed: (a) lowering the pH toward 6·7 favors gelation; (b) deoxyhemoglobin S gels more readily in 6 mm-2,3-diphosphoglycerate than in its total absence; (c) 1 m-NaCl and l m-KCl inhibit gelation. The known favoring of gelation by warming is confirmed by the equilibrium method and is about 2% change in minimal gelling concentration per degree. The effects of pH and high ionic strengths are consistent with contributions of specific polar interactions to gel structure. The effect of 2,3-diphosphoglycerate probably depends on known structural changes which this cofactor induces rather than on alteration of the allosteric quaternary structure equilibrium.

Original languageEnglish (US)
Pages (from-to)445-452,IN1-IN2,453-458
JournalJournal of Molecular Biology
Volume80
Issue number3
DOIs
StatePublished - Nov 5 1973

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Effects of pH, 2,3-diphosphoglycerate and salts on gelation of sickle cell deoxyhemoglobin'. Together they form a unique fingerprint.

  • Cite this