Linking Conformation Change to Hemoglobin Activation Via Chain-Selective Time-resolved Resonance Raman Spectroscopy on Protoheme/Mesoheme Hybrids

Time-resolved Resonance Raman spectra are reported for Hb tetramers, in which the αand β chains are selectively substituted with mesoheme. The Soret absorption band shift in meso- relative to protoheme permits chain-selective excitation of heme RR spectra. The evolution of these spectra following HbCO photolysis show that geminate recombination rates and yields are the same for the two chains, consistent with recent results on 15N-heme isotopomer hybrids. The spectra also reveal systematic shifts in the deoxy-heme ν4 and νFe-His) RR bands, which are anti-correlated. These shifts are resolved for the successive intermediates in the protein structure, which have previously been determined from time-resolved UVRR spectra. Both chains show Fe-His bond compression in the immediate photoproduct, which relaxes during the formation of the first intermediate, Rdeoxy (0.07 μs), in which the proximal F-helix is proposed to move away from the heme. Subsequently, the Fe-His bond weakens, more so for the α than the β chains. The weakening is gradual for the β chains, but abrupt for the α chains, coinciding with completion of the R-T quaternary transition, at 20μs. Since the transition from fast- to slow-rebinding Hb also occurs at 20μs, the drop in the α chain νFe-His supports the localization of ligation restraint to tension in the Fe-His bond, at least in the α-chains. The mechanism is more complex in the β chains

Subunit-Selective Interrogation of CO Recombination in Carbonmonoxy Hemoglobin by Isotope-Edited Time-Resolved Resonance Raman Spectroscopy

Hemoglobin (Hb) is an allosteric tetrameric protein made up of αβ heterodimers. The α and β chains are similar, but are chemically and structurally distinct. To investigate dynamical differences between the chains, we have prepared tetramers in which the chains are isotopically distinguishable, via reconstitution with 15N-heme. Ligand recombination and heme structural evolution, following HbCO dissociation, was monitored with chain selectivity by resonance Raman (RR) spectroscopy. For α but not for β chains, the frequency of the ν4 porphyrin breathing mode increased on the microsecond time scale. This increase is a manifestation of proximal tension in the Hb T-state, and its time course is parallel to the formation of T contacts, as determined previously by UVRR spectroscopy. Despite the localization of proximal constraint in the α chains, geminate recombination was found to be equally probable in the two chains, with yields of 39 ± 2%. We discuss the possibility that this equivalence is coincidental, in the sense that it arises from the evolutionary pressure for cooperativity, or that it reflects mechanical coupling across the αβ interface, evidence for which has emerged from UVRR studies of site mutants

Deformations of calibrated subbundles of Euclidean spaces via twisting by special sections

We extend the "bundle constructions" of calibrated submanifolds, due to Harvey--Lawson in the special Lagrangian case, and to Ionel--Karigiannis--Min-Oo in the cases of exceptional calibrations, by "twisting" the bundles by a special (harmonic, holomorphic, parallel) section of a complementary bundle. The existence of such deformations shows that the moduli space of calibrated deformations of these "calibrated subbundles" includes deformations which destroy the linear structure of the fibre.Comment: 16 pages, no figures. Version 2: Only minor cosmetic and typographical revisions. To appear in "Annals of Global Analysis and Geometry.

An investigation into the feasibility of myoglobin-based single-electron transistors

Myoglobin single-electron transistors were investigated using nanometer- gap platinum electrodes fabricated by electromigration at cryogenic temperatures. Apomyoglobin (myoglobin without heme group) was used as a reference. The results suggest single electron transport is mediated by resonant tunneling with the electronic and vibrational levels of the heme group in a single protein. They also represent a proof-of-principle that proteins with redox centers across nanometer-gap electrodes can be utilized to fabricate single-electron transistors. The protein orientation and conformation may significantly affect the conductance of these devices. Future improvements in device reproducibility and yield will require control of these factors