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Peptide and protein mapping by 252Cf-plasma desorption mass spectrometry
A Tsarbopoulos1, G W Becker, J L Occolowitz
1Department of Pharmacology, Mayo Clinic, Rochester, Minnesota 55905.
Analytical Biochemistry
|May 15, 1988
Summary
Plasma desorption mass spectrometry (PDMS) offers a fast and sensitive method for peptide mapping, verifying protein sequences with minimal sample. This technique aids in confirming DNA-deduced sequences of proteins like interleukin-2.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- Peptide mapping using mass spectrometry is a standard method for protein sequence verification.
- Fast atom bombardment mass spectrometry (FAB-MS) is a well-established technique for this purpose.
Purpose of the Study:
- To describe and evaluate a novel peptide mapping approach using 252Cf-plasma desorption mass spectrometry (PDMS).
- To demonstrate the utility of PD mapping for screening DNA-deduced protein sequences.
Main Methods:
- Proteins (recombinant interleukin-2 and human growth hormone) were chemically or enzymatically cleaved.
- Peptide molecular weights were determined using 252Cf-PDMS.
- Mass spectrometric data was compared with predicted peptide masses from DNA sequences.
Main Results:
- PD mapping successfully screened DNA-deduced sequences of recombinant interleukin-2 and human growth hormone.
- The non-destructive nature of PDMS allowed for further peptide characterization via modification.
- The method proved to be fast and sensitive, requiring low nanomole quantities.
Conclusions:
- PD mapping using 252Cf-PDMS is a viable and efficient alternative for protein sequence verification.
- The technique facilitates rapid confirmation of protein sequences derived from DNA.
- PDMS offers a sensitive and versatile tool for peptide analysis in molecular biology and biochemistry.