Proteome-wide identification of metamorphic protein candidates using mass spectrometry.

ABSTRACT: How protein folds remodel at low temperature remains poorly mapped at the proteome scale. We combine brief, temperature-controlled proteolysis with LC-MS/MS to chart cold-dependent conformational changes in Escherichia coli lysate. Methodological validation with the metamorphic protein Sa1V90T-whose α/β-plait and 3α states interchange between 30 °C and 5 °C-showed that a high-protease, short-digestion regime detects temperature-dependent exposure at four peptides, whereas the temperature-insensitive variant Sa1V90TV52D is largely unchanged. Applied to lysate, the workflow identifies >750 proteolytic sites across >250 abundant proteins (>10% of the identified proteome) whose local susceptibility shifts as temperature decreases from 35 °C to 5 °C, including five Sa1V90T peptides recap

SUBMITTER: Ouan B 

PROVIDER: S-EPMC12642558 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature