title: Liganded hemoglobin structural perturbations by the allosteric effector L35.
abstract: Effector binding to liganded hemoglobin (Hb) provides a new understanding of structural determinants of Hb function. L35, a bezafibrate-related compound, is one of the more potent synthetic regulators of Hb oxygen (O(2)) affinity. In the presence of inositol hexaphosphate and bezafibrate (or derivatives), liganded Hb at low pH (pH approximately 6.5) exhibits extremely low O(2) affinity and very low cooperativity. In this study, the nature of L35 binding to COHbA at pH 6.35, an altered R-state, is presented. Solution-active site-specific spectroscopic probings by front-face fluorescence and circular dichroism reveal that L35 induces a global heterogeneous conformation in COHbA at pH 6.35 that includes: a T-like structural feature at the alpha1beta2 interface; an R-like structural feature within the heme environment; and an intermediate-like state at the central cavity. These long-range structural perturbations appear to stem from L35 binding to two classes of binding sites: the central cavity (primarily at the alphaalpha cleft) and the surface. These results indicate that L35 induces an allosteric transition species, characterized by domain-specific tertiary and quaternary-like conformation within a global R-quaternary structure.
authors.affiliation: Department of Medicine, Division of Hematology, Montefiore Medical Center, Bronx, New York, USA.
authors.forename: Qiuying
authors.initials: Q
authors.lastname: Chen
authors.forename: Iraj
authors.initials: I
authors.lastname: Lalezari
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Nagel
authors.forename: Rhoda Elison
authors.initials: RE
authors.lastname: Hirsch
mesh_terms: D001665:Binding Sites; D006454:Hemoglobins / Q000032:analysis / Q000737:chemistry*; D008024:Ligands; D010671:Phenylurea Compounds / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
chemical_list: D006454:Hemoglobins; D008024:Ligands; D010671:Phenylurea Compounds; C064070:L 35