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Group Members Research Interests
Prof. Paul N. Barlow
Professor of Structural Biology
Our research focus is on protein structure.
The main tool we use is solution state NMR, but we also employ
calorimetry, circular dichroism, hydrodynamics, molecular modelling,
mass spectrometry and fluorescence spectroscopy to complement
the high-resolution structure information.
For example, we are interested in multi-modular proteins such
as are found in the complement system. Many of these are large,
extended, flexible and glycosylated. We use NMR to study small
fragments of two or three modules, calorimetry to study module-module
interactions and a combination of hydrodynamics and molecular
modelling to extrapolate to bigger fragments or complete proteins.
Dr. Dusan Uhrin
Lecturer in NMR spectroscopy
My research deals with the application of NMR to interesting biological problems as well as with the development of new techniques for biomolecular NMR spectroscopy.
At the heart of my research are the studies of protein-glycosaminoglycan interactions. We are investigating several polyanion binding sites of factor H, a crucial regulator of the alternative pathway of complement. In another study we are characterising interaction of heparan and dermatan sulfates with NK1, an isoform of the
hepatocyte growth factor/scatter factor. Understanding these interactions on a molecular level can lead to rational design of drugs and vaccines.
I am developing new NMR techniques for the conformational analysis of carbohydrates, focusing in particular on the methods for the measurement of residual dipolar coupling constants and long-range carbon-carbon coupling constants. We are also developing new NMR techniques for the solution structure determination of protein-GAG complexes.
Dr. Janice Bramham
Lecturer in Structural Biology
I am interested in the three-dimensional structures and intrinsic
dynamics of biological macromolecules with the ultimate aim of
understanding their structure-function relationships. My current work
is focused upon proteins of the human complement system, which plays an
essential role in our immune response to infection.
I am determining tertiary structures and investigating specific protein:protein interactions that occur
in macromolecular assemblies. The principal technique I employ is high-resolution, multi-dimensional
NMR spectroscopy which I complement with other biophysical techniques, such as mass spectrometry
and isothermal titration calorimetry.
Dr. Juraj Bella
email: juraj.bella@ed.ac.uk
NMR Facilities Manager
Dr. Andrew Herbert
email: a.herbert@ed.ac.uk
Dr. Graeme Ball
email: graeme.ball@ed.ac.uk
Dr. David Kavanagh
email: davidkavanagh@doctors.org.uk
Dr. Marie Phelan
email: marie.phelan@ed.ac.uk
The membrane attack complex (MAC) is a key component at the terminus of the comple-
ment pathways; it has the ability to kill cells by puncturing the plasma membrane resulting in
cytolysis. Initiated by the proteolytic cleavage of C5, the subsequent formation of the
MAC is a non-catalytic, self-assembly process whereby five soluble proteins rapidly
organise themselves into a membrane-penetrating macromolecular complex. My research,
utilising high-field NMR spectroscopy, aims to provide an insight into the tertiary
structures of the MAC proteins. I am investigating the protein-protein interactions and
conformational changes involved in the MAC assembly process and in the maintenance of the
stable macromolecular complex. This work, developed and coordinated by Dr Janice Bramham,
is funded by the MRC.
Bärbel Blaum
email: baerbel.blaum@ed.ac.uk
Glycosaminoglycan-Protein interaction are thought to be at the centre of important
processes of host recognition within the human Complement System. My PhD work focuses on
the purification and characterization of small heparin-derived oligosaccharides and
subsequent binding studies to Complement Factor H modules. I aim to characterize the GAG-protein interaction on a molecular level using mostly NMR based techniques. In particular, Paramagnetic Relaxation Enhancements generated by spin-labeled oligosaccharides prepared by our collaborators.
Chris Fenton
email: chris.fenton@ed.ac.uk
I am working on a solution structure of a factor H module belonging to one of its three
heparin binding sites. Factor H is an immune system regulator of the complement system.
It works by preventing the alternative pathway of the complement system becoming active on
host cell membranes, which would otherwise lead to cell lysis. To recognise host from non-host
cell membranes, factor H binds to polyanions found on the surface of host cells.
Our collaborators have confirmed, by Gel Mobility Shift Assay, that our recombinant module
does bind to a fully sulfated heparin-derived tetrasaccharide. This finding broadens the
scope of my PhD program to include the characterisation of the interaction between fH and heparin
sulphate/heparin tetrasaccharides using NMR.
Henry Hocking
email: H.G.Hocking@sms.ed.ac.uk
My PhD project focuses on the regulatory region of the human complement
regulator factor H. This region, located at the N-terminus of factor H,
is made up of four CCP modules and is responsible for accelerating the
decay of the C3 and C5 convertases of the alternative pathway as well
as acting as a cofactor to the proteolytic degradation of C3b by factor
I. These two modes of action provide protection of host cells from
non-specific attack by the alternative pathway of complement.
My main objective is to solve the solution structure of overlapping
module pairs (~14 kDa per pair) within this regulatory region in order
to reconstruct the entire functional site. My current work involves
cloning, expression, purification of the different module pairs
followed by their structure determination using NMR spectroscopy
methods. Triple module constructs are also being studied to validate
this approach and to further assess the inter-modular orientations and
dynamics within this region.
Lan Jin
email: L.Jin@sms.ed.ac.uk
I study the gas phase structures of heparin-derived oligosaccharides by
using ion mobility mass spectrometry and molecular modelling.
In solution I use various techniques of NMR spectroscopy to study a
conformation of a fully sulfated heparin-derived tetrasaccharide.
In our efforts to characterize the
conformation of glycosidic linkages of carbohydrates I am working on
the development of techniques for the measurement of long-range
carbon-carbon coupling constants in natural abundance 13C samples.
Further work is in progress on the development of robust methods for
the measurement of proton-proton residual dipolar coupling constants of
carbohydrates.
Marc Lenoir
email: m.lenoir@sms.ed.ac.uk
Tumors growth is often associated with neovascularisation events which
can be detected via specific markers during the earliest stages of the
cancer progression. It has been shown that the integrin family
potentially represents one of these markers. General Electric
Health has designed a series of cyclic peptides which are aimed at
binding specifically integrin avbIII and by carrying a radio labelling
metal are detectable by imaging techniques. Our goal is to study the
three dimensional structure of these peptides by using NMR
spectroscopy.
Isabell Pechtl
email: I.C.Pechtl@sms.ed.ac.uk
For my PhD project I am employing a combination of fluorescence resonance energy transfer (FRET) and NMR to study the link between conformation and function in factor H. Factor H is composed of 20 CCP modules (~1200 amino acids). Factor H regulates the amplification of the alternative complement pathway and sequence variations in factor H (both mutations and SNPs) are linked to several diseases of the kidneys and eye.
My goal is to use FRET (fluorescence resonance energy transfer) to measure long-range interatomic distances in factor H and thereby support NMR-based structural investigations that rely on short-range (NOE) restraints. I shall also use paramagnetic NMR to obtain further structural restraints.
By using site-directed mutagenesis, cysteine residues can be introduced into the CCP modules of factor H at strategic points and subsequently be used for attachment of fluorescent tags such as caged lanthanides that have the added advantage of being paramagnetic.
Elin Pless
email: elin.pless@ed.ac.uk
My PhD project involves the GABAB receptor, which is a membrane protein involved in synaptic transmission
in various brain regions and forms the basis of a powerful and versatile inhibitory system in the mammalian brain.
The N-terminal region of the GABABR1a consists of two complement protein modules (CCP-modules),
which are highly conserved protein sequences. The CCP-modules seem to function as binding sites
for macromolecules. The main aim of my project is to investigate possible interactions between
the GABABR1a CCP modules and various proteins. This is explored by using the Yeast Two Hybrid
system (YTH), Biacore and different protein pull-down methods as well as functional studies
in the model system Xenopus Laevis.
Patience Tetteh-Quarcoo
email: elin.pless@ed.ac.uk
Christoph Schmidt
email:s0460219@sms.ed.ac.uk
My PhD project deals with the module-module orientation and structure of the central CCP
modules (12 to 14) from the fluid phase regulator of the alternative pathway - factor H.
Distinct module orientation/interactions are likely to originate from the diverse CCP
module sequences and the variable type and numbers of residues within the intervening
linkers. This is impossible, at present, to model and therefore there is a particular
interest in the experimental determination of module-module orientations.
The use of paramagnetic probe molecules in the field of NMR has become a powerful
technique with which to refine solution structures. It is valuable for obtaining
long-range restrains, up to a maximum distance of 40 Å. It is therefore particularly
useful for refining the structure of extended loops, or obtaining intermodular orientations
within multidomain proteins where few NOEs occur between modules.
One aim of this project is to apply this technique to determine the relative orientation
of the central CCP modules (12 to 14) of factor H.
Dinesh Soares
email: D.C.Soares@sms.ed.ac.uk
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