Biology subjects

Tame, J. R. H.

Publications and source records attributed to Tame, J. R. H..

3 recordsLinked to original sources

bioRxiv2025

Conformational analysis of liganded human hemoglobin by cryo electron microscopy

The long-standing debate on the preferred conformation of liganded hemoglobin (Hb) in solution has yet to be completely resolved. While some studies have used lyophilized human hemoglobin for structural studies by cryo-EM, we recently presented the first cryo-EM analysis of freshly prepared human and crocodilian Hbs. Further three-dimensional (3D) classification analysis of these datasets reveals distinct structural characteristics. CO-bound adult human Hb (CO-HbA) shows a mixture of conformations, with the R2 conformation most populated, R strongly represented, and other intermediate states present in sufficient quantity to produce maps. CO-bound crocodile Hb showed the R conformation and, unexpectedly, a smaller population of molecules in a T-like conformation. The amino acid substitution Glu {beta}39 [->] Arg, unique to crocodilian Hbs, appears to favour the R conformation over R2.

molecular biology↗

bioRxiv2021

Crystal sturctures of Scone, pseudosymmetric folding of a symmetric designer protein

Recent years have seen a raise in the development of computational proteins including symmetric ones. We recently developed a nine-fold symmetric {beta}-propeller protein named Cake. Here we wanted to further engineer this protein to a three-fold symmetric nine-bladed propeller using computational design. Two nine-bladed propeller proteins were designed, named Scone-E and Scone-R. Crystallography however revealed the structure of both designs to adopt an eight-fold conformation with distorted termini, leading to a pseudo-symmetric protein. One of the proteins could only be crystallized upon addition of a polyoxometalate highlighting the usefulness of these molecules as a crystallisation additive.

systems biology↗

bioRxiv2020

Shape and size complementarity induced formation of supramolecular protein assemblies with metal-oxo clusters

The controlled formation of protein supramolecular assemblies is challenging but it could provide an important route for the development of hybrid biomaterials. In this work, we demonstrate formation of well-defined complexes formed between the 8-fold symmetrical designer protein Tako8 and soluble metal-oxo clusters from the family of Anderson-Evans, Keggin and ZrIV- substituted Wells-Dawson polyoxometalates. A combination of x-ray crystallography and solution studies showed that metal-oxo clusters are able to serve as linker nodes for the bottom-up creation of protein based supramolecular assemblies. Our findings indicate that clusters with larger size and negative charge are capable of modulating the crystal packing of the protein, highlighting the need for a size and shape complementarity with the protein node for optimal alteration of the crystalline self-assembly.

biochemistry↗