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Principles of Client Enrichment in Multicomponent Biomolecular Condensates

Biomolecular condensates are commonly organized by a small number of scaffold molecules that drive phase separation together with client molecules that do not condense on their own but become selectively recruited into the dense phase. A central open question is how client recruitment feeds back on scaffold interactions to determine condensate composition. Here we address this problem in a reconstituted focal adhesion system composed of focal adhesion kinase (FAK) and phosphorylated p130Cas (Cas) as scaffolds and the adaptor protein paxillin (PXN) as a client. We show that both FAK phosphorylation and PXN recruitment produce a common compositional response in which FAK becomes enriched while Cas is depleted within the condensate. To interpret these observations, we develop two complementary theoretical descriptions. First, within a two-component Flory-Huggins framework, we show that phosphorylation can be captured by either strengthening heterotypic FAK-Cas interactions or increasing the effective number of interaction-relevant segments on FAK, both of which bias partitioning toward FAK-rich condensates. Second, we introduce a minimal three-component Flory-Huggins theory without an explicit solvent and map it onto an effective two-component description, demonstrating that client recruitment renormalizes homotypic and heterotypic scaffold interactions. Analytical predictions for the location of the critical point are tested in reconstituted multicomponent systems through PXN addition, showing that client recruitment alone tunes proximity to criticality and reshapes condensate composition. Together, our results reveal distinct yet convergent physical routes by which post-translational modification and client recruitment control scaffold composition in multicomponent condensates.