The binding of small diatomic ligands such as carbon monoxide or dioxygen to heme proteins is among the simplest biological processes known. Still, it has taken many decades to understand the mechanistic aspects of th...The binding of small diatomic ligands such as carbon monoxide or dioxygen to heme proteins is among the simplest biological processes known. Still, it has taken many decades to understand the mechanistic aspects of this process in full detail. Here, we compare ligand binding in three heme proteins of the globin family, myoglobin, a dimeric hemoglobin, and neuroglobin. The combination of structural, spectroscopic, and kinetic experiments over many years by many laboratories has revealed common properties of globins and a clear mechanistic picture of ligand binding at the molecular level. In addition to the ligand binding site at the heme iron, a primary ligand docking site exists that ensures efficient ligand binding to and release from the heme iron. Additional, secondary docking sites can greatly facilitate ligand escape after its dissociation from the heme. Although there is only indirect evidence at present, a preformed histidine gate appears to exist that allows ligand entry to and exit from the active site. The importance of these features can be assessed by studies involving modified proteins（via site-directed mutagenesis） and comparison with heme proteins not belonging to the globin family.展开更多
In recent years,multi-photon 3D laser printing has become a widely used tool for the fabrication of micro-and nanostructures for a large variety of applications.Typically,thorough sample characterisation is key for an...In recent years,multi-photon 3D laser printing has become a widely used tool for the fabrication of micro-and nanostructures for a large variety of applications.Typically,thorough sample characterisation is key for an efficient optimisation of the printing process.To date,three-dimensional microscopic inspection has usually been carried out on finished 3D printed microstructures,that is,using ex-situ approaches.In contrast,in-situ 3D characterization tools are desirable for quickly assessing the quality and properties of 3D printed microstructures.Along these lines,we present and characterise a Fourier-domain optical coherence tomography(FD-OCT)system that can be readily integrated into an existing 3D laser lithography setup.We demonstrate its capabilities by examining different 3D printed polymer microstructures immersed in a liquid photoresist.In such samples,local reflectivity arises from the(refractive-index)contrasts between the polymerised and non-polymerised regions.Thus,the refractive index of the printed material can be extracted.Furthermore,we demonstrate that the reflectivity of polymer-monomer transitions exhibits time-dependent behaviour after printing.Supported by transfer-matrix calculations,we explain this effect in terms of the time-dependent graded-index transition originating from monomer diffusion into the polymer matrix.Finally,we show exemplary 3D reconstructions of printed structures that can be readily compared with 3D computer designs.展开更多
基金supported by the Deutsche Forschungsgemeinschaft (DFG, grant Ni291/10)
文摘The binding of small diatomic ligands such as carbon monoxide or dioxygen to heme proteins is among the simplest biological processes known. Still, it has taken many decades to understand the mechanistic aspects of this process in full detail. Here, we compare ligand binding in three heme proteins of the globin family, myoglobin, a dimeric hemoglobin, and neuroglobin. The combination of structural, spectroscopic, and kinetic experiments over many years by many laboratories has revealed common properties of globins and a clear mechanistic picture of ligand binding at the molecular level. In addition to the ligand binding site at the heme iron, a primary ligand docking site exists that ensures efficient ligand binding to and release from the heme iron. Additional, secondary docking sites can greatly facilitate ligand escape after its dissociation from the heme. Although there is only indirect evidence at present, a preformed histidine gate appears to exist that allows ligand entry to and exit from the active site. The importance of these features can be assessed by studies involving modified proteins（via site-directed mutagenesis） and comparison with heme proteins not belonging to the globin family.
基金This work was funded by the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under Germany’s Excellence Strategy 2082/1-390761711(Excellence Cluster“3D Matter Made to Order”).
文摘In recent years,multi-photon 3D laser printing has become a widely used tool for the fabrication of micro-and nanostructures for a large variety of applications.Typically,thorough sample characterisation is key for an efficient optimisation of the printing process.To date,three-dimensional microscopic inspection has usually been carried out on finished 3D printed microstructures,that is,using ex-situ approaches.In contrast,in-situ 3D characterization tools are desirable for quickly assessing the quality and properties of 3D printed microstructures.Along these lines,we present and characterise a Fourier-domain optical coherence tomography(FD-OCT)system that can be readily integrated into an existing 3D laser lithography setup.We demonstrate its capabilities by examining different 3D printed polymer microstructures immersed in a liquid photoresist.In such samples,local reflectivity arises from the(refractive-index)contrasts between the polymerised and non-polymerised regions.Thus,the refractive index of the printed material can be extracted.Furthermore,we demonstrate that the reflectivity of polymer-monomer transitions exhibits time-dependent behaviour after printing.Supported by transfer-matrix calculations,we explain this effect in terms of the time-dependent graded-index transition originating from monomer diffusion into the polymer matrix.Finally,we show exemplary 3D reconstructions of printed structures that can be readily compared with 3D computer designs.
