When Less is MORE: Counterintuitive Stoichiometries and Cellular Abundances are Essential for 2 ABC transporters’ function

ABSTRACT: Prokaryotes acquire essential nutrients primarily through ATP-binding cassette (ABC) importers, 18 consisting of an ATPase, a permease, and a substrate-binding protein. These importers are highly 19 underrepresented in proteomic databases, limiting our knowledge about their cellular copy numbers,20 component stoichiometry, and the mechanistic implications of these parameters. We developed a tailored 21 proteomic approach to compile the most comprehensive dataset to date of the E. coli 'ABC importome'. 22 Functional assays and analysis of deletion strains revealed mechanistic features linking molecular 23 mechanisms to cellular abundances, co-localization, and component stoichiometries. We observed 4-5 24 orders of magnitude variation in import system abundances, with copy numbers tuned to nutrient hierarchies 25 essential for growth. Abundances of substrate-binding proteins are unrelated to their substrate binding 26 affinities but are tightly, yet inversely, correlated with their interaction affinity with permeases. 27 Counterintuitive component stoichiometries are crucial for function, offering insights into the design 28 principles of multi-component protein systems, potentially extending beyond ABC importers.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli

SUBMITTER: Tamar Ziv  

LAB HEAD: Oded Lewinson

PROVIDER: PXD061770 | Pride | 2025-05-26

REPOSITORIES: Pride