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Michal Or-Guil

Publications and source records attributed to Michal Or-Guil.

4 recordsLinked to original sources
PubMed2005

SPOT synthesis: reliability of array-based measurement of peptide binding affinity.

Peptide arrays prepared by the SPOT synthesis technology have emerged as a proteomic tool to study molecular recognition and identify biologically active peptides. However, it was previously not clear how accurately signal intensities obtained by probing peptide arrays for protein binding really reflect the dissociation constants of the protein-peptide complexes. Using the monoclonal antibody CB4-1 as a model system, we systematically compared dissociation constants of antibody-peptide complexes with signal intensities obtained using the SPOT technology. By analyzing a set of peptides possessing different affinities to the antibody, we determined the strengths of the SPOT screening method. The accuracy of the measured results was improved by taking regional trends in the membrane surface into account. A model based on the mass action law compares well with the experimental results. Interestingly, the applied concentrations of the binding partners do not directly correspond to the effective concentrations in the assay. We show that the SPOT technology is an accurate method for assigning the spots' measured signal intensities to three different binding affinity classes. The dissociation constants of the intermediate region were found to be between pK(dis)=5 and pK(dis)=7. Altering the experimental parameters causes a directed change of this region.

Antibodies, Monoclonal↗
PubMed2005

A mathematical model of protein degradation by the proteasome.

The proteasome is the major protease for intracellular protein degradation. The influx rate of protein substrates and the exit rate of the fragments/products are regulated by the size of the axial channels. Opening the channels is known to increase the overall degradation rate and to change the length distribution of fragments. We develop a mathematical model with a flux that depends on the gate size and a phenomenological cleavage mechanism. The model has Michaelis-Menten kinetics with a V(max) that is inversely related to the length of the substrate, as observed in the in vitro experiments. We study the distribution of fragment lengths assuming that proteasomal cleavage takes place at a preferred distance from the ends of a protein fragment, and find multipeaked fragment length distributions similar to those found experimentally. Opening the gates in the model increases the degradation rate, increases the average length of the fragments, and increases the peak in the distribution around a length of 8-10 amino acids. This behavior is also observed in immunoproteasomes equipped with PA28. Finally, we study the effect of re-entry of processed fragments in the degradation kinetics and conclude that re-entry is only expected to affect the cleavage dynamics when short fragments enter the proteasome much faster than the original substrate. In summary, the model proposed in this study captures the known characteristics of proteasomal degradation, and can therefore help to quantify MHC class I antigen processing and presentation.

Antigen Presentation↗
PubMed2002

Drifting pattern domains in a reaction-diffusion system with nonlocal coupling.

Drifting pattern domains (DPDs), i.e., moving localized patches of traveling waves embedded in a stationary (Turing) pattern background and vice versa, are observed in simulations of a reaction-diffusion model with nonlocal coupling. Within this model, a region of bistability between Turing patterns and traveling waves arises from a codimension-2 Turing-wave bifurcation (TWB). DPDs are found within that region in a substantial distance from the TWB. We investigated the dynamics of single interfaces between Turing and wave patterns. It is found that DPDs exist due to a locking of the interface velocities, which is imposed by the absence of space-time defects near these interfaces.

Journal Article↗