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FT-ICR-MS approach to monitor asparagine deamidation and its isomers products in collagen from ancient bones
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Perez Hurtado, Pilar (2013) FT-ICR-MS approach to monitor asparagine deamidation and its isomers products in collagen from ancient bones. PhD thesis, University of Warwick.
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WRAP_THESIS_Perez Hurtado_2013.pdf - Submitted Version Download (6Mb) | Preview |
Official URL: http://webcat.warwick.ac.uk/record=b2689072~S1
Abstract
This project investigates the use of ultrahigh resolution mass spectrometry along with
fragmentation techniques such as electron capture dissociation (ECD) and
collisionally activated dissociation (CAD) to study deamidation of ancient bone, by
using potential deamidation markers present in bovine collagen standards. With the
application of these techniques, several marker peptides present in the digested
protein standard of bovine collagen were successfully assigned. The sequences of
these peptides correlated well with the reported sequences for bovine collagen in the
literature.
FT-ICR-MS was used to monitor deamidation of collagen by following a shift of
+0.948 Da in the spectrum, resulting in a mass difference of 19 mDa from the 13C of
the non-deamidated form and the 12C of the deamidated form, which can be difficult
to assign due to overlap with the 13C isotopic distribution in peptides. The rate
constants for the deamidation reaction were calculated, and the extent of deamidation
before sample handling was determined. The methodology developed was then
applied to collagen extracted from real bone samples, both modern and ancient,
proving to be a useful method for monitoring asparagine deamidation before sample
preparation. Differentiation of the isomers products of deamidation (aspartic and
isoaspartic acid) were successfully assigned (where possible) using the diagnostic
ions originated from their ECD spectra.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QP Physiology | ||||
Library of Congress Subject Headings (LCSH): | Ion cyclotron resonance spectrometry, Fourier transform spectroscopy, Deamination, Collagen | ||||
Official Date: | July 2013 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | Department of Chemistry | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | O’Connor, Peter B. | ||||
Sponsors: | Engineering and Physical Sciences Research Council (EPSRC) | ||||
Extent: | xxi, 205 leaves : illustrations. | ||||
Language: | eng |
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