title: Oxygenation properties of human erythrocytes containing exclusively alpha-nitrosyl hemoglobin: a promising blood transfusant candidate.
abstract: We have prepared human erythrocytes that contain exclusively alpha-nitrosyl hemoglobin (Hb), i.e., alpha(Fe-NO)2beta(Fe-O2)2, by incorporating nitric oxide (NO) into erythrocytes in a well-controlled nitrosylation process. The amount of alpha(Fe-NO) corresponding to 50% of the total heme content of the erythrocytes and exclusive binding of NO to alpha-subunits of intraerythrocytic Hb were confirmed by EPR. Oxygenation experiments on the intraerythrocytic alpha-nitrosyl Hb over a wide range of pH showed that: (1) the oxygen affinity of cell-free and intraerythrocytic alpha-nitrosyl Hbs were much lower than native Hb in their respective environments; (2) the oxygenation characteristics of the intraerythrocytic alpha-nitrosyl Hb in the acidic range was similar to that of the cell-free alpha-nitrosyl Hb in the presence of 2,3-diphosphoglycerate; and (3) the apparent Bohr effect in the intraerythrocytic alpha-nitrosyl Hb was dramatically diminished. This can be due to a restricted variation in intraerythrocytic pH in the alkaline region and the presence and/or production of endogenous 2,3-diphosphoglycerate. By comparing oxygen saturation characteristics, it was found that the intraerythrocytic alpha-nitrosyl Hb, despite its halved oxygen carrying capacity, could deliver more oxygen than DPG-depleted erythrocytes under similar experimental conditions. This makes alpha-nitrosyl Hb-containing erythrocytes a promising candidate for blood transfusant.
authors.affiliation: Department of Biochemistry and Biophysics and The Johnson Foundation, University of Pennsylvania Medical Center, Philadelphia 19104-6059, USA. ants@mail.med.upenn.edu
authors.forename: A
authors.initials: A
authors.lastname: Tsuneshige
authors.forename: T
authors.initials: T
authors.lastname: Yonetani
mesh_terms: D001802:Blood Substitutes / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D004912:Erythrocytes / Q000737:chemistry* / Q000378:metabolism; D006454:Hemoglobins / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism*; D010108:Oxyhemoglobins
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D010108:Oxyhemoglobins; C008657:nitrosyl hemoglobin; D010100:Oxygen