In situ structural studies of membrane protein megacomplexes

Membrane protein complexes are essential for cellular functions, which rely on both constituent protein structures and their interactions within native membranes. While in vitro methods have successfully yielded high-resolution structures of individual proteins and subcomplexes, these approaches typically require detergent extraction and extensive purification, which can disrupt the native membrane environment and potentially alter the supramolecular organization. In situ structural biology has therefore emerged as an effective strategy to overcome these limitations by directly visualizing macromolecular machines within their physiological context. With continuous technological advancements, several recent studies have resolved in situ structures of large protein complexes at high or even near-atomic resolution. This review focuses on recent in situ high-resolution studies of membrane protein megacomplexes, highlighting key technical innovations, structural insights, and the remaining challenges and opportunities in the field.