Structure of deoxyhemoglobin: ionizable groups and polyethylene glycol.
X-ray studies on deoxy-hemoglobin have been reported on crystals grown under conditions of high (about 2.5 M) and low salt (about 0.1 M). The high-salt crystals were grown in water, whereas the low-salt crystals were prepared in polyethylene glycol solutions (m.w. 6000-8000 Da, 10-30% w/v). Oxygen-binding characteristics of hemoglobin in these two environments differ radically. In salt solutions, hemoglobin binds oxygen with a p50 value of about 5 torr of oxygen and in a cooperative manner characterized with an n value of Hill varying from two to three. In polyethylene glycol solutions, hemoglobin crystals are oxygenated with a p50 value of about 270 torr of oxygen, without exhibiting a cooperative effect. I report on a detailed study of the X-ray data defining the dimer interface (alpha1beta2) of these two forms of hemoglobin. The study reveals that the main difference between the two structures lies in the number and arrangement of the water molecules and in distances between ionizable side chains in the dimer interface. I propose that these differences lead to significant shifts in the pK values of the ionizable groups in the dimer interface.
Saroff
Harry A
HA
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA. saroff@helix.nih.gov
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019281:Dimerization; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D006859:Hydrogen / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism*; D011487:Protein Conformation
D006454:Hemoglobins; D011092:Polyethylene Glycols; D006859:Hydrogen; C100003:deoxyhemoglobin; D010100:Oxygen