Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Molecular engineering of a polymer of tetrameric hemoglobins
Autore:
Fronticelli, C; Arosio, D; Bobofchak, KM; Vasquez, GB;
Indirizzi:
Univ Maryland, Dept Biochem & Mol Biol, Sch Med, Baltimore, MD 21201 USA Univ Maryland Baltimore MD USA 21201 ol, Sch Med, Baltimore, MD 21201 USA Johns Hopkins Univ, Sch Med, Dept Anesthesiol, Baltimore, MD USA Johns Hopkins Univ Baltimore MD USA Dept Anesthesiol, Baltimore, MD USA Ctr Adv Res Biotechnol, Rockville, MD 20850 USA Ctr Adv Res Biotechnol Rockville MD USA 20850 ol, Rockville, MD 20850 USA Natl Inst Stand & Technol, Rockville, MD USA Natl Inst Stand & Technol Rockville MD USA & Technol, Rockville, MD USA
Titolo Testata:
PROTEINS-STRUCTURE FUNCTION AND GENETICS
fascicolo: 3, volume: 44, anno: 2001,
pagine: 212 - 222
SICI:
0887-3585(20010815)44:3<212:MEOAPO>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
RECOMBINANT HUMAN HEMOGLOBIN; CROSS-LINKED HEMOGLOBINS; BLOOD SUBSTITUTE; CELL; TRANSFUSION;
Keywords:
recombinant hemoglobins; polymerization; genetic engineering; dynamic light scattering; autoxidation; thermal stability; heme pocket; oxygen affinity;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Fronticelli, C Univ Maryland, Dept Biochem & Mol Biol, Sch Med, Baltimore,MD 21201 USA Univ Maryland Baltimore MD USA 21201 ltimore, MD 21201 USA
Citazione:
C. Fronticelli et al., "Molecular engineering of a polymer of tetrameric hemoglobins", PROTEINS, 44(3), 2001, pp. 212-222

Abstract

We have engineered a recombinant mutant human hemoglobin, Hb Prisca beta (S9C+C93A+C112G), which assembles in a polymeric form. The polymerization isobtained through the formation of intermolecular S-S bonds between cysteine residues introduced at position beta9, on the model of Hb Porto Alegre (beta 9Ser --> Cys) (Bonaventura and Riggs, Science 1967;155:800-802). C beta93 and C beta 112 were replaced in order to prevent formation of spurious S-S bonds during the expression, assembly, and polymerization events. Dynamic light scattering measurements indicate that the final polymerization product is mainly formed by 6 to 8 tetrameric hemoglobin molecules. The samplepolydispersity Q = 0.07 +/- 0.02, is similar to that of purified human hemoglobin (Q = 0.02 +/- 0.02), consistent with a good degree of homogeneity. In the presence of strong reducing agents, the polymer reverts to its tetrameric form. During the depolymerization process, a direct correlation is observed between the hydrodynamic radius and the light scattering of the system, which, in turn, is proportional to the mass of the protein. We interpret this to indicate that the hemoglobin molecules are tightly packed in the polymer with no empty spaces. The tight packing of the hemoglobin moleculessuggests that the polymer has a globular shape and, thus, allows estimation of its radius. An illustration of an arrangement of a finite number of tetrameric hemoglobin molecules is presented. The conformational and functional characteristics of this polymer, such as heme pocket conformation, stability to denaturation, autoxidation rate, oxygen affinity, and cooperativity, remain similar to those of tetrameric human hemoglobin. (C) 2001 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 08/04/20 alle ore 11:53:42