dc.contributor.advisor | Lindberg, Iris | |
dc.contributor.author | Miller, Larissa K. | |
dc.date.accessioned | 2011-08-08T13:47:41Z | |
dc.date.available | 2011-08-08T13:47:41Z | |
dc.date.issued | 1996 | |
dc.identifier.uri | http://hdl.handle.net/10920/23115 | |
dc.description | viii, 38 p. | en_US |
dc.description.abstract | The substrate and inhibitory specificity of PC2, a mammalian enzyme involved in
the cleavage of many constitutively expressed protein precursors, was studied
using substrate phage display and combinatorial chemistry. In the method of
substrate phage display, a multitude of substrate sequences are displayed as fusion
proteins on filamentous phage particles. The cleaved phage are propagated and
submitted to additional rounds of protease selection to further enrich for good
substrates. DNA sequencing of the cleaved phage is used to identify the preferred
substrate sequences. To modify substrate phage display technique using a low pH
enzyme the method was optimized for elution with a low pH buffer. After 6
rounds of sorting a substrate phage library containing S random amino acids
(XXXXX) there was no consensus sequence among the eluted phage.
Combinatorial· chemistry using a positional scanning synthetic combinational
library (PS-SCL) offers a unique and rapid approach for the identification of
individual active compounds from libraries made up of millions of compounds.
The PS-SCL described here is made up of six individual positional peptide
libraries, each one consisting of hexamers with a single position defined and five
positions as mixtures. The PS-SCL aided in the identification of two inhibitory
sequences for PC2. The first is Ac-IIRVKR-NH2 with an ICSO value of 13 µM
and the second is Ac-IKRVKR-NH2 with an 1C50 value of 36 µM. These two
sequences show the recognition sequence for PC2 to be possibly -RVKR-. While
PC2 and PC 1 are in the same family and are both neuroendocrine processing
enzymes they do not share similar inhibitory or simulatory poly-amino acid
sequences. Modifications of the substrate phage display method for selection with
a low pH enzyme are still required. The inhibitory sequence found through using
a PS-SCL further aids in understanding the specificity and active site of the
enzyme PC2. | en_US |
dc.description.sponsorship | Department of Biochemistry. Louisiana State University Medical Center. Louisiana State University. New Orleans, Louisiana. | |
dc.format.mimetype | application/pdf | |
dc.language.iso | en_US | en_US |
dc.publisher | Kalamazoo College | en_US |
dc.relation.ispartof | Kalamazoo College Biology Senior Individualized Projects Collection | |
dc.relation.ispartofseries | Senior Individualized Projects. Biology; | |
dc.rights | U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder. | |
dc.title | A Survey of Prohormone Convertase 2 (PC2) Substrate Specificity Using Substrate Phage Display and Combinatorial Chemistry | en_US |
dc.type | Thesis | en_US |
KCollege.Access.Contact | If you are not a current Kalamazoo College student, faculty, or staff member, email dspace@kzoo.edu to request access to this thesis. | |