{Reference Type}: Journal Article {Title}: Experimental and computational approaches for membrane protein insertion and topology determination. {Author}: Duart G;GraƱa-Montes R;Pastor-Cantizano N;Mingarro I; {Journal}: Methods {Volume}: 226 {Issue}: 0 {Year}: 2024 Jun 9 {Factor}: 4.647 {DOI}: 10.1016/j.ymeth.2024.03.012 {Abstract}: Membrane proteins play pivotal roles in a wide array of cellular processes and constitute approximately a quarter of the protein-coding genes across all organisms. Despite their ubiquity and biological significance, our understanding of these proteins remains notably less comprehensive compared to their soluble counterparts. This disparity in knowledge can be attributed, in part, to the inherent challenges associated with employing specialized techniques for the investigation of membrane protein insertion and topology. This review will center on a discussion of molecular biology methodologies and computational prediction tools designed to elucidate the insertion and topology of helical membrane proteins.