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Studies on peptidases

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Tunnicliffe, Alan, 1963- (1990) Studies on peptidases. PhD thesis, University of Warwick.

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Official URL: http://webcat.warwick.ac.uk/record=b3226542~S15

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Abstract

This study details the methods of purification of the peptidase enzyme, Diaminopeptidase I (DAPI, Cathepsin C) and an acetyl-amino acid releasing enzyme called Acetylaminopeptidase (AcAP). The preparations obtained were fully characterized and identified as being homogenous preparations of the above enzymes. Together with Diaminopeptidase IV (DAPIV) it is intended to use these enzymes to hydrolyse unknown derivatized peptides and proteins for sequencing studies.

Several different techniques were used in my studies. They included three different methods for the estimation of molecular weights, two methods for the determination of isoelectric points, an activity stain, inhibition studies, substrate specificity, studies and storage studies.

Chapter 3 details the use of DAPI in an immobilized form to hydrolyze a peptide, with subsequent separation of the digestion products with sequence analysis by HPLC.

This work culminated in a publication (Ref70).
Many results confirm those found for the enzymes by previous workers. However, the metal ion inhibition studies of AcAP, in particular, contains some results contrary to some published results but possible reasons for this are offered.

In summary, this study details methods for the purification of two enzymes postulated to be of use in the sequencing of some derivatized peptides. Further work, which takes the form of the use of the enzyme cocktail was carried out but these studies were incomplete and therefore not included in this study. However the techniques which I would have been used and the conditions which would have been employed have been included in the introduction for the sake of completeness.

Item Type: Thesis or Dissertation (PhD)
Subjects: Q Science > QP Physiology
Library of Congress Subject Headings (LCSH): Peptidase
Official Date: March 1990
Dates:
DateEvent
March 1990UNSPECIFIED
Institution: University of Warwick
Theses Department: Department of Chemistry
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Hutchinson, D. W. (David Wesley)
Format of File: pdf
Extent: [16], 146, [4] leaves : illustrations
Language: eng

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