TIE-UP-SIN: a novel method for enhanced identification of protein-protein interactions.

ABSTRACT: Proteins function through complex interaction networks that govern nearly all aspects of cellular physiology. Identifying protein-protein interactions (PPIs) under native conditions remains challenging due to the transient nature of many complexes and technical limitations of conventional approaches. We present TIE-UP-SIN (Targeted Interactome Experiment for Unknown Proteins by Stable Isotope Normalization), a robust and reproducible method for in vivo identification of PPIs. This approach combines metabolic labeling with ¹⁵N isotopes, reversible in vivo formaldehyde crosslinking, affinity purification, and quantitative mass spectrometry. TIE-UP-SIN is specifically designed to preserve transient or weak interactions during purification and to quantify interaction p

SUBMITTER: Schedlowski M 

PROVIDER: S-EPMC12434083 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature