Laboratory of Molecular Biophysics Laboratory Journal 2000 Dr. M. E. M. Noble

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Martin Noble

Protein kinases, SH2-SH3 domains and arylamine N-acetyl transferase

Overview

Work in this group addresses structure function relationships of medically important proteins from two different areas: proteins involved in regulatory processes inside eukaryotic cells, and the enzyme arylamine N-acetyltransferase. Research in these areas involves a combination of practical and theoretical approaches. Practically, proteins are studied by functional and biophysical characterisation, particularly using isothermal titration calorimetry in collaboration with the group of John Ladbury at University College London, and by structure determination through X-ray crystallography. On the theoretical side, we have been developing tools to aid in the analysis of properties of protein surfaces.

This year has seen the completion of a study into the interaction between focal adhesion kinase and fyn, a member of the Src family of non-receptor tyrosine kinases. This study has been part of a long standing collaboration with the group of Professor Iain Campbell into the fyn/FAK system. Combining results from this group with work carried out in Professor Campbell's group has allowed us to see the effects on the structure and dynamics of the regulatory fragment of a Src-family kinase, when it forms an interaction key to downstream signalling from focal adhesions. We have also succeeded in crystallising a novel domain of FAK, the focal adhesion kinase domain, and we hope to study its structure and interactions

We have also broadened the scope of our research into the interactions of the -chain of the T-cell receptor (TCR). TCR interactions play a major role in the development of an immune response, primarily through a signalling pathway involving the non-receptor tyrosine kinase ZAP 70. We have developed purification schemes for ZAP 70 expressed in insect cells, involving either immobilised metal affinity chromatography, or an affinity column which exploits the affinity of ZAP 70 for its natural ligands, the immunoreceptor tyrosine-based activation motifs (ITAMs). We have also started to explore the interaction of ITAMs with the HIV protein NEF. This interaction may be important in the progression of HIV infections through down-regulation of the host immune response.

Development of theoretical methods has centred on increasing the capabilities of the program Aesop. Aesop is an interactive molecular illustration program which is designed to allow novel analyses of the character of protein surfaces, and to prepare output in the form of moving as well as static images. This year, Aesop has been given a graphical user interface which takes advantage of the GTK toolkit, and it has been extended to interface smoothly with GRID for quantitative and qualitative analysis of the chemical character of protein surfaces. Analysis of surface conservation has also been extended to allow sophisticated analyses of conservation and differences among sub-families within an aligned set of sequences.

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