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Titolo:
Polymer structure and solubility of deoxyhemoglobin S in the presence of high concentrations of volume-excluding 70-kDa dextran - Effects of non-S hemoglobins and inhibitors
Autore:
Bookchin, RM; Balazs, T;
Indirizzi:
Albert Einstein Coll Med, Dept Med, Bronx, NY 10461 USA Albert Einstein Coll Med Bronx NY USA 10461 Dept Med, Bronx, NY 10461 USA Univ Chicago, Dept Mol Genet & Cell Biol, Chicago, IL 60637 USA Univ Chicago Chicago IL USA 60637 enet & Cell Biol, Chicago, IL 60637 USA Univ Cambridge, Physiol Lab, Cambridge CB2 3EG, England Univ Cambridge Cambridge England CB2 3EG Lab, Cambridge CB2 3EG, England
Titolo Testata:
JOURNAL OF BIOLOGICAL CHEMISTRY
fascicolo: 10, volume: 274, anno: 1999,
pagine: 6689 - 6697
SICI:
0021-9258(19990305)274:10<6689:PSASOD>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
SICKLE-CELL HEMOGLOBIN; CRYSTALLIZATION; MICROMETHOD; MUTANT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Bookchin, RM Albert Einstein Coll Med, Dept Med, 1300 Morris Pk Ave, Bronx, NY 10461 USA Albert Einstein Coll Med 1300 Morris Pk Ave Bronx NY USA 10461
Citazione:
R.M. Bookchin e T. Balazs, "Polymer structure and solubility of deoxyhemoglobin S in the presence of high concentrations of volume-excluding 70-kDa dextran - Effects of non-S hemoglobins and inhibitors", J BIOL CHEM, 274(10), 1999, pp. 6689-6697

Abstract

Earlier observations indicated that volume exclusion by admired non-hemoglobin macromolecules lowered the polymer solubility ("C-sat") of deoxyhemoglobin (Hb) S, presumably by increasing its activity. In view of the potential usefulness of these observations for in vitro studies of sickling-relatedpolymerization, we examined the ultrastructure, solubility behavior, and phase distributions of deoxygenated mixtures of Hb S with 70-kDa dextran, a relatively inert, low ionic strength space-filling macromolecule. Increasing admixture of dextran progressively lowered the C-sat of deoxyHb S, With 12 g/dl dextran, a 5-fold decrease in apparent C-sat ("dextran-C-sat") was obtained together with acceptable sensitivity and proportionality with the standard C-sat when assessing the effects of non-S Hb admixtures (A, C, and F) or polymerization inhibitors (alkylureas or phenylalanine). The volume fraction of dextran excluding Hb was 70-75% of total deoxy Hb-dextran (12 g/dl) volumes. Electron microscopy showed polymer fibers and fiber-to-crystaltransitions indistinguishable from those formed without dextran, Thus whenHb quantities are limited, as with genetically engineered recombinant Hbs or transgenic sickle mice, the dextran-C-sat provides convenient and reliable screening of effects of Hb S modifications on polymerization under near-physiological conditions, avoiding problems of high ionic strength.

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Documento generato il 21/09/20 alle ore 06:25:54