Molecular and genetic analysis of domains mediating laminin self interactions

• Applicant: Professor Dr. Mats Paulsson, since 2/2005
• Subject Area: Dermatology
• Term: from 2000 to 2008
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5250700

Laminins form an organised network within the basement membrane in part by virtue of their potential to self aggregate. This aggregation occurs due to the interaction of their three N-terminal domains (domain VI) and laminins lacking one or more of these domains do not aggregate in vitro. We will recombinantly express and purify the domain VI from the murine laminin b1 chain. The domain will be tested for its ability to interact with both intact laminin-1 and with N-terminal domains from other chains. To allow the elucidation of its interaction surface domain VI will be crystallised and its spatical structure determined. Substitution of possibly significant residues with alanines by site directed mutagenesis, followed by determination of laminin-1 affinity, will be used to define a binding surface. Where significant alteration in binding is seen, the mutation will be repeated in the genome of murine ES cells which will then be used to produce embryoid bodies and mice. In these in vivo systems the biological consequences of abolished laminin self interactions will be determined.

• DFG Programme: Priority Programmes
• Subproject of: SPP 1086: Genetic and Molecular Analysis of Basement Membranes and Basement Membrane Anchorage
• Ehemaliger Antragsteller: Dr. Neil R. Smyth, until 2/2005