title: Studying macromolecular complex stoichiometries by peptide-based mass spectrometry.
abstract: A majority of cellular functions are carried out by macromolecular complexes. A host of biochemical and spectroscopic methods exists to characterize especially protein/protein complexes, however there has been a lack of a universal method to determine protein stoichiometries. Peptide-based MS, especially as a complementary method to the MS analysis of intact protein complexes, has now been developed to a point where it can be employed to assay protein stoichiometries in a routine manner. While the experimental demands are still significant, peptide-based MS has been successfully applied to analyze stoichiometries for a variety of protein complexes from very different biological backgrounds. In this review, we discuss the requirements especially for targeted MS acquisition strategies to be used in this context, with a special focus on the interconnected experimental aspects of sample preparation, protein digestion, and peptide stability. In addition, different strategies for the introduction of quantitative peptide standards and their suitability for different scenarios are compared.
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute for Biophysical Chemistry, Goettingen, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Wohlgemuth
authors.forename: Christof
authors.initials: C
authors.lastname: Lenz
authors.forename: Henning
authors.initials: H
authors.lastname: Urlaub
mesh_terms: D046911:Macromolecular Substances / Q000032:analysis / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D040901:Proteomics
chemical_list: D046911:Macromolecular Substances; D010455:Peptides
keywords: Mass spectrometry; Protein complex; SRM; Stoichiometry; Technology