Electrospray Ionization Mass Spectrometry on Hydrophobic Peptides Electroeluted from Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Application to the Topology of the Sarcoplasmic Reticulum Ca2+ ATPase
✍ Scribed by M. Lemaire; S. Deschamps; J.V. Moller; J.P. Lecaer; J. Rossier
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 628 KB
- Volume
- 214
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
We describe a method to prepare proteins and peptides in a state suitable for exact determination of molecular mass by electrospray ionization mass spectrometry after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroelution. The utility of the procedure, in conjunction with (\mathrm{N})-terminal sequencing, in defining the C-terminal end of the peptide fragments produced by proteolysis of sarcoplasmic reticulum (\mathrm{Ca}^{2+}) ATPase with V8 is demonstrated. The application of mass spectrometry aids significantly the use of proteolytic enzymes for topological studies of membrane proteins, and SDS-PAGE is preferable to reverse-phase HPLC for separation of membraneous, hydrophobic peptides and proteins. of 1993 Academic Press, Inc.