title: Time-resolved fluorescence of hemoglobin species.
abstract: We used time-resolved fluorescence in the pico- to nanosecond time range to monitor the presence of tetramers, dimers and monomers in carbonmonoxyhemoglobin (COHb) solutions and to investigate how their distributions change under different experimental conditions. Comparison of fluorescence lifetime computed from the atomic coordinates of COHb (Vasquez et al., 1996) with those experimentally measured allowed identification of molecular species present in the hemoglobin solution. It was possible to observe modification of the distribution of tetramers, dimers, monomers and species with disordered hemes produced by different experimental conditions. Protein concentration affected the detectable lifetimes, indicating increasing amounts of dimers and monomers at low protein concentrations, while the amount of inverted hemes was not modified. Titration with up to 1 M NaCl modified only the extent of dissociation of hemoglobin into dimers, without affecting heme inversion and monomer formation. Hyperbaric pressure increased the amounts of dimers and monomers. This is the first time that monomeric subunits of hemoglobin have been detected at neutral pH in the normal system.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Maryland Medical School, Baltimore 21201, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Gryczynski
authors.forename: S
authors.initials: S
authors.lastname: Beretta
authors.forename: J
authors.initials: J
authors.lastname: Lubkowski
authors.forename: A
authors.initials: A
authors.lastname: Razynska
authors.forename: I
authors.initials: I
authors.lastname: Gryczynski
authors.forename: E
authors.initials: E
authors.lastname: Bucci
mesh_terms: D002263:Carboxyhemoglobin / Q000737:chemistry*; D006801:Humans; D009994:Osmolar Concentration; D011312:Pressure; D013050:Spectrometry, Fluorescence
chemical_list: D002263:Carboxyhemoglobin