This project aims at elucidating the pathophysiology of hemolytic anemias secondary to defects in red cell metabolism. We will investigate in detail the glycolytic process of the red cell """"""""in vitro"""""""" utilizing newly developed techniques that allow metabolic flow to proceed unimpeded by metabolic block. This is a novel approach to the study of red cell metabolism, that shall permit analysis of glycolysis in detail, in conditions similar to those that exist in the red cell """"""""in vivo"""""""". We will investigate in detail the biochemical changes that occur during the process of red cell aging """"""""in vivo"""""""". We will examine, through analysis of glycolytic intermediates in age-fractionated red cells, the progressive development of metabolic alterations. We will study in great detail two key enzymes of glycolysis - hexokinase and pyruvate kinase. We will attempt to clarify their molecular structure and to assess the role played by isozymic shifts in the process of molecular aging of enzymes, versus the possibility that this may result from post-translational modifications.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
Project #
Application #
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Columbia University (N.Y.)
Schools of Medicine
New York
United States
Zip Code
Murakami, K; Kanno, H; Miwa, S et al. (1999) Human HKR isozyme: organization of the hexokinase I gene, the erythroid-specific promoter, and transcription initiation site. Mol Genet Metab 67:118-30
Wang, X; Piomelli, S; Peacocke, M et al. (1997) Erythropoietic protoporphyria: four novel frameshift mutations in the ferrochelatase gene. J Invest Dermatol 109:688-91
Murakami, K; Piomelli, S (1997) Identification of the cDNA for human red blood cell-specific hexokinase isozyme. Blood 89:762-6
Wang, X (1996) Molecular characterization of a novel defect occurring de novo associated with erythropoietic protoporphyria. Biochim Biophys Acta 1316:149-52
Ostasiewicz, L T; Huang, J L; Wang, X et al. (1995) Human protoporphyria: genetic heterogeneity at the ferrochelatase locus. Photodermatol Photoimmunol Photomed 11:18-21
Wang, X; Poh-Fitzpatrick, M; Chen, T et al. (1995) Systematic screening for RNA with skipped exons--splicing mutations of the ferrochelatase gene. Biochim Biophys Acta 1271:358-62
Wang, X; Poh-Fitzpatrick, M; Taketani, S et al. (1994) Screening for ferrochelatase mutations: molecular heterogeneity of erythropoietic protoporphyria. Biochim Biophys Acta 1225:187-90
Wang, X; Poh-Fitzpatrick, M; Piomelli, S (1994) A novel splicing mutation in the ferrochelatase gene responsible for erythropoietic protoporphyria. Biochim Biophys Acta 1227:25-7
Wang, X; Poh-Fitzpatrick, M; Carriero, D et al. (1993) A novel mutation in erythropoietic protoporphyria: an aberrant ferrochelatase mRNA caused by exon skipping during RNA splicing. Biochim Biophys Acta 1181:198-200
Tilton, W M; Seaman, C; Piomelli, S (1993) Measurement of radiolabeled lactate production using lactate monooxygenase. Anal Biochem 208:400-2

Showing the most recent 10 out of 11 publications