New light on allostery: dynamic resonance Raman spectroscopy of hemoglobin kempsey.
On the basis of static and time-resolved resonance Raman spectroscopy of HbA and of a mutant, HbK (Dalpha99N), a specific reaction coordinate is proposed for the allosteric transition in human hemoglobin. The heme is held between proximal (F) and distal (E) helices, whose orientation is responsive to forces generated by ligation and deligation. The E and F helices are in turn tethered via H-bonds to the A and H helices. These outer helices follow the E-F motion, thereby repositioning the N- and C-termini, which form the intersubunit salt bridges in the T quaternary structure. When the T state interface is weakened by Asp --> Asn substitution at a quaternary H-bond (HbK), the Fe-His bond is relaxed and becomes responsive to allosteric effectors. The same E-F motion is observed in HbK, but the A-H following motion is delayed, relative to HbA, as is the Asn H-bond formation.
Hu
X
X
Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA.
Rodgers
K R
KR
Mukerji
I
I
Spiro
T G
TG
D000494:Allosteric Regulation; D006418:Heme / Q000737:chemistry; D006455:Hemoglobins, Abnormal / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012399:Rotation; D013059:Spectrum Analysis, Raman
D006455:Hemoglobins, Abnormal; C032577:hemoglobin Kempsey; D006418:Heme