Roles of hemoglobin allostery in hypoxia-induced metabolic alterations in erythrocytes

Simulation and its verification by metabolome analysis

Ayako Kinoshita, Kosuke Tsukada, Tomoyoshi Soga, Takako Hishiki, Yuki Ueno, Yoichi Nakayama, Masaru Tomita, Makoto Suematsu

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

When erythrocytes are exposed to hypoxia, hemoglobin (Hb) stabilizes in theT-state by capturing 2,3-bisphosphoglycerate. This process could reduce the intracellular pool of glycolytic substrates, jeopardizing cellular energetics. Recent observations suggest that hypoxia-induced activation of glycolytic enzymes is correlated with their release from Band III (BIII) on the cell membrane. Based on these data, we developed a mathematical model of erythrocyte metabolism and compared hypoxia-induced differences in predicted activities of the enzymes, their products, and cellular energetics between models with and without the interaction of Hb with BIII. The models predicted that the allostery-dependent Hb interaction with BIII accelerates consumption of upstream glycolytic substrates such as glucose 6-phosphate and increases downstream products such as phosphoenolpyruvate. This prediction was consistent with metabolomic data from capillary electrophoresis mass spectrometry. The hypoxia-induced alterations in the metabolites resulted from acceleration of glycolysis, as judged by increased conversion of [13C]glucose to [13C]lactate. The allostery-dependent interaction of Hb with BIII appeared to contribute not only to maintenance of energy charge but also to further synthesis of 2,3-bisphosphoglycerate, which could help sustain stabilization of T-state Hb during hypoxia. Furthermore, such an activation of glycolysis was not observed when Hb was stabilized in R-state by treating the cells with CO. These results suggest that Hb allostery in erythrocytes serves as an O2-sensing trigger that drives glycolytic acceleration to stabilize intracellular energetics and promote the ability to release O 2 from the cells.

Original languageEnglish
Pages (from-to)10731-10741
Number of pages11
JournalJournal of Biological Chemistry
Volume282
Issue number14
DOIs
Publication statusPublished - 2007 Apr 6

Fingerprint

Metabolome
Hemoglobins
Erythrocytes
2,3-Diphosphoglycerate
Glycolysis
Chemical activation
Capillary electrophoresis
Glucose-6-Phosphate
Phosphoenolpyruvate
Enzyme Activation
Metabolomics
Capillary Electrophoresis
Substrates
Enzymes
Cell membranes
Carbon Monoxide
Metabolites
Metabolism
Mass spectrometry
Hypoxia

ASJC Scopus subject areas

  • Biochemistry

Cite this

Roles of hemoglobin allostery in hypoxia-induced metabolic alterations in erythrocytes : Simulation and its verification by metabolome analysis. / Kinoshita, Ayako; Tsukada, Kosuke; Soga, Tomoyoshi; Hishiki, Takako; Ueno, Yuki; Nakayama, Yoichi; Tomita, Masaru; Suematsu, Makoto.

In: Journal of Biological Chemistry, Vol. 282, No. 14, 06.04.2007, p. 10731-10741.

Research output: Contribution to journalArticle

@article{59a3a2b71aa648b8a749977e16503fd3,
title = "Roles of hemoglobin allostery in hypoxia-induced metabolic alterations in erythrocytes: Simulation and its verification by metabolome analysis",
abstract = "When erythrocytes are exposed to hypoxia, hemoglobin (Hb) stabilizes in theT-state by capturing 2,3-bisphosphoglycerate. This process could reduce the intracellular pool of glycolytic substrates, jeopardizing cellular energetics. Recent observations suggest that hypoxia-induced activation of glycolytic enzymes is correlated with their release from Band III (BIII) on the cell membrane. Based on these data, we developed a mathematical model of erythrocyte metabolism and compared hypoxia-induced differences in predicted activities of the enzymes, their products, and cellular energetics between models with and without the interaction of Hb with BIII. The models predicted that the allostery-dependent Hb interaction with BIII accelerates consumption of upstream glycolytic substrates such as glucose 6-phosphate and increases downstream products such as phosphoenolpyruvate. This prediction was consistent with metabolomic data from capillary electrophoresis mass spectrometry. The hypoxia-induced alterations in the metabolites resulted from acceleration of glycolysis, as judged by increased conversion of [13C]glucose to [13C]lactate. The allostery-dependent interaction of Hb with BIII appeared to contribute not only to maintenance of energy charge but also to further synthesis of 2,3-bisphosphoglycerate, which could help sustain stabilization of T-state Hb during hypoxia. Furthermore, such an activation of glycolysis was not observed when Hb was stabilized in R-state by treating the cells with CO. These results suggest that Hb allostery in erythrocytes serves as an O2-sensing trigger that drives glycolytic acceleration to stabilize intracellular energetics and promote the ability to release O 2 from the cells.",
author = "Ayako Kinoshita and Kosuke Tsukada and Tomoyoshi Soga and Takako Hishiki and Yuki Ueno and Yoichi Nakayama and Masaru Tomita and Makoto Suematsu",
year = "2007",
month = "4",
day = "6",
doi = "10.1074/jbc.M610717200",
language = "English",
volume = "282",
pages = "10731--10741",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - Roles of hemoglobin allostery in hypoxia-induced metabolic alterations in erythrocytes

T2 - Simulation and its verification by metabolome analysis

AU - Kinoshita, Ayako

AU - Tsukada, Kosuke

AU - Soga, Tomoyoshi

AU - Hishiki, Takako

AU - Ueno, Yuki

AU - Nakayama, Yoichi

AU - Tomita, Masaru

AU - Suematsu, Makoto

PY - 2007/4/6

Y1 - 2007/4/6

N2 - When erythrocytes are exposed to hypoxia, hemoglobin (Hb) stabilizes in theT-state by capturing 2,3-bisphosphoglycerate. This process could reduce the intracellular pool of glycolytic substrates, jeopardizing cellular energetics. Recent observations suggest that hypoxia-induced activation of glycolytic enzymes is correlated with their release from Band III (BIII) on the cell membrane. Based on these data, we developed a mathematical model of erythrocyte metabolism and compared hypoxia-induced differences in predicted activities of the enzymes, their products, and cellular energetics between models with and without the interaction of Hb with BIII. The models predicted that the allostery-dependent Hb interaction with BIII accelerates consumption of upstream glycolytic substrates such as glucose 6-phosphate and increases downstream products such as phosphoenolpyruvate. This prediction was consistent with metabolomic data from capillary electrophoresis mass spectrometry. The hypoxia-induced alterations in the metabolites resulted from acceleration of glycolysis, as judged by increased conversion of [13C]glucose to [13C]lactate. The allostery-dependent interaction of Hb with BIII appeared to contribute not only to maintenance of energy charge but also to further synthesis of 2,3-bisphosphoglycerate, which could help sustain stabilization of T-state Hb during hypoxia. Furthermore, such an activation of glycolysis was not observed when Hb was stabilized in R-state by treating the cells with CO. These results suggest that Hb allostery in erythrocytes serves as an O2-sensing trigger that drives glycolytic acceleration to stabilize intracellular energetics and promote the ability to release O 2 from the cells.

AB - When erythrocytes are exposed to hypoxia, hemoglobin (Hb) stabilizes in theT-state by capturing 2,3-bisphosphoglycerate. This process could reduce the intracellular pool of glycolytic substrates, jeopardizing cellular energetics. Recent observations suggest that hypoxia-induced activation of glycolytic enzymes is correlated with their release from Band III (BIII) on the cell membrane. Based on these data, we developed a mathematical model of erythrocyte metabolism and compared hypoxia-induced differences in predicted activities of the enzymes, their products, and cellular energetics between models with and without the interaction of Hb with BIII. The models predicted that the allostery-dependent Hb interaction with BIII accelerates consumption of upstream glycolytic substrates such as glucose 6-phosphate and increases downstream products such as phosphoenolpyruvate. This prediction was consistent with metabolomic data from capillary electrophoresis mass spectrometry. The hypoxia-induced alterations in the metabolites resulted from acceleration of glycolysis, as judged by increased conversion of [13C]glucose to [13C]lactate. The allostery-dependent interaction of Hb with BIII appeared to contribute not only to maintenance of energy charge but also to further synthesis of 2,3-bisphosphoglycerate, which could help sustain stabilization of T-state Hb during hypoxia. Furthermore, such an activation of glycolysis was not observed when Hb was stabilized in R-state by treating the cells with CO. These results suggest that Hb allostery in erythrocytes serves as an O2-sensing trigger that drives glycolytic acceleration to stabilize intracellular energetics and promote the ability to release O 2 from the cells.

UR - http://www.scopus.com/inward/record.url?scp=34249859307&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34249859307&partnerID=8YFLogxK

U2 - 10.1074/jbc.M610717200

DO - 10.1074/jbc.M610717200

M3 - Article

VL - 282

SP - 10731

EP - 10741

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -