Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Ashley Colemon - Hampton University
Co-Author(s): Isi Ero-Tolliver, Hampton University, Hampton,VA
Assembly and integrity of collagen IV networks is essential for the filtering function of the kidney glomeruli. Disruption of this network which occurs in diseases such as diabetic nephropathy, Goodpasture (GP) syndrome and Alport syndrome can lead to reduced kidney function and ultimately to renal failure. Using high resolution NMR and mass spectrometry, a novel sulfilimine bond (S=N) between methionine and hyroxylysine in the NC1 domains of collagen IV was identified as important for stabilization of these networks. This was the first report of a sulfilimine bond in a biomolecule and it appears to be in the basement membranes of animal species ranging from cnidarians to humans suggesting it plays a fundamental role in tissue function and it is known to play a role in the pathogenesis of GP syndrome. As little is known about the formation of this novel bond, we began to elucidate the mechanism of sulfilimine bond formation. Recently, we identified peroxidasin as the multi-domain peroxidase responsible for catalyzing the sulfilimine bond formation by using its peroxidase and n-terminus of the immunoglobulin domain. Our current work focuses on investigating and identifying the critical amino acids of peroxidasin responsible for catalyzing sulfilimine bond formation. We are biologically engineering a variety of truncated constructs that contain specific regions of the peroxidasin. We anticipate that our results will provide insight into amino acids necessary and sufficient for catalysis of sulfilimine bond formation and enhancing crosslinking of the basement membranes.
Funder Acknowledgement(s): NSF ( National Science Foundation) Louis Stokes Alliance for Minority Participation (LSAMP).
Faculty Advisor: Isi Ero-Tolliver, Isi.firstname.lastname@example.org
Role: I am in the beginning stages of the research. My role involves critical analysis of the Peroxidasin sequence to identify regions for mutation and truncation. Using current literature and deductive reasoning, I will be selecting mutants of priority for immediate cloning. I am in the process of setting up the laboratory (ordering cell lines, reagents, and additional research equipment) in order to proceed with the research that was initially performed at Vanderbilt University. With my Principal Investigator (PI) and I's continued collaboration with PIs at Vanderbilt University, we are establishing a site for cloning and protein purification here at Hampton University.