Crystal structure of deoxygenated hemoglobin in complex with sphingosine phosphate and 2,3-Bisphosphoglycerate

2,3-Bisphosphoglyceric acid; 2,3-bisphosphoglycerate; 2,3-BPG; 2,3-diphosphoglyceric acid; 2,3-diphosphoglycerate; 2,3-DPG; (2~{R})-2,3-diphosphonooxypropanoic acid

(2S,3R,4E)-2-amino-3-hydroxyoctadec-4-en-1-yl dihydrogen phosphate

Structural and Functional Insight of Sphingosine 1-Phosphate-Mediated Pathogenic Metabolic Reprogramming in Sickle Cell Disease.

VLSPADKTNVKAAWGKVGAHAGEYGAEALERMFLSFPTTKTYFPHFDLSHGSAQVKGHGKKVADALTNAVAHVDDMPNAL
SALSDLHAHKLRVDPVNFKLLSHCLLVTLAAHLPAEFTPAVHASLDKFLASVSTVLTSKYR

VHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLGAFSDGLAHLDN
LKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVANALAHKYH

0.2 M sodium acetate trihydrate, 0.1 M sodium cacodylate trihydrate, pH 6.5 and 30% PEG 8000

National Institutes of Health/National Institute on Minority Health and Health Disparities (NIH/NIMHD)

National Institutes of Health/National Cancer Institute (NIH/NCI)

Author provided the following evidence of direct S1P binding to hemoglobin: S1P beads successfully pulled down HbA from normal human erythrocyte lysates, while sphingosine or lysophosphatidic acid beads failed to pull down Hb. In addition, co-binding of 2,3-DPG and S1P to hemoglobin (Hb) is required for S1P-induced decrease in Hb-O2 affinity. We measured Oxygen Equilibration Curves (OEC) of purified HbA in the presence of 2,3-DPG along with S1P. We found that S1P decreased HbA-O2 binding affinity with increased P50 in a dose-dependent manner. Thus, we provide biochemical and functional evidence that S1P binds directly to Hb but requires cobinding of 2,3-DPG to decrease the protein affinity for O2

Hemoglobin, Sphingosine-1-phosphate, Complex, ALLOSTERIC EFFECTOR, OXYGEN TRANSPORT