Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.14/13153
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- Title
- Evaluation of chemical derivatisation methods for protein identification using MALDI MS/MS
- Related
- International journal of peptide research and therapeuti, Vol. 12, Issue 3, p.225-235
- DOI
- 10.1007/s10989-006-9026-3
- Publisher
- Springer Verlag
- Date
- 2006
- Author/Creator
- Joss, Janice L
- Author/Creator
- Molloy, Mark P
- Author/Creator
- Hinds, Lyn A
- Author/Creator
- Deane, Elizabeth M
- Description
- In proteomic studies, assigning protein identity from organisms whose genomes are yet to be completely sequenced remains a challenging task. For these organisms, protein identification is typically based on cross species matching of amino acid sequence obtained from collision induced dissociation (CID) of peptides using mass spectrometry. The most direct approach of "de novo" sequencing is slow and often difficult, due to the complexity of the resultant CID spectra. For MALDI-MS, this problem has been addressed by using chemical derivatisation to direct peptide fragmentation, thereby simplifying CID spectra and facilitating "de novo" interpretation. In this study, milk whey proteins from the tammar wallaby (“Macropus eugenii”) were used to evaluate three chemical derivatisation methods compatible with MALDI MS/MS. These methods included (i) guanidination and sulfonation using chemically-assisted fragmentation (CAF), (ii) guanidination and sulfonation using 4-sulfophenyl isothiocyanate (SPITC) and (iii) derivatising the epsilon-amino group of lysine residues with Lys Tag 4H. Derivatisation with CAF and SPITC resulted in more protein identification than Lys Tag 4H. Sulfonation using SPITC was the preferred method due to the low cost per experiment, the reactivity with both lysine and arginine terminated peptides and the resultant simplified MS/MS spectra.
- Description
- 11 page(s)
- Subject Keyword
- cross species matching
- Subject Keyword
- "de novo" sequencing
- Subject Keyword
- MALDI MS/MS
- Subject Keyword
- guanidination
- Subject Keyword
- sulfonation
- Resource Type
- journal article
- Organisation
- Macquarie University. Dept. of Chemistry and Biomolecular Sciences
- Organisation
- Macquarie University. Division of Environmental and Life Sciences
- Identifier
- http://hdl.handle.net/1959.14/13153
- Identifier
- ISSN:1573-3904
- Identifier
- mq-rm-2006005823
- Language
- eng
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