Date of Award
Chemistry and Biochemistry
CC BY-NC-ND 4.0
Recently, a novel Kunitz inhibitor of trypsin-like enzymes has been discovered in bovine platelets that has a higher degree of association for plasmin than trypsin, and thereby has been termed bovine platelet plasmin inhibitor (BPPI). Basic problems in the purification, resulting in protein that was essentially insoluble, has hampered structural determinations by x-ray or NMR analysis. An investigation into the cause of the protein insolubility has revealed that an excessively high concentration of acetonitrile, used to elute the protein from a C18 reversed-phase cartridge, was most probably the cause of the insolubility. Concentrated BPPI solutions permitted the preparation of a suitable sample for structural determination by NMR, and preliminary $\sp1$H-NOESY and $\sp1$H-TOCSY spectra have been obtained. However, given the high degree of sequence homology with a bovine pancreatic trypsin inhibitor (BPTI), a protein whose three-dimensional structure has been extensively characterized, a structural determination of BPPI has been performed using molecular modelling. Circular dichroism studies have indicated that the secondary structural elements in BPPI are similar to those of BPPI and are accurately predicted in the model. A search for a similar protein in human platelets has resulted in the discovery of a 12,000 Da (by SDS-PAGE) protein that also inhibits trypsin and plasmin and has preliminarily been termed human platelet plasmin inhibitor (HPPI). (Abstract shortened by UMI.)Dept. of Chemistry and Biochemistry. Paper copy at Leddy Library: Theses & Major Papers - Basement, West Bldg. / Call Number: Thesis1995 .B34. Source: Masters Abstracts International, Volume: 34-06, page: 2373. Adviser: Lana Lee. Thesis (M.Sc.)--University of Windsor (Canada), 1995.
Baldwin, Jeffrey Stephen., "Purification, characterization, and functional investigations of novel platelet proteins." (1995). Electronic Theses and Dissertations. 3245.