Dataset Information

The structural dynamics of full length divisome transmembrane proteins FtsQ, FtsB, and FtsL in FtsQBL complex formation

ABSTRACT: Bacterial divisome is a group of proteins governing cell division. Out of the 15 critical proteins in Escherichia coli, the transmembrane protein complex FtsQBL plays an essential role in regulating the action. Although extensive efforts have been made, there is limited information regarding the interaction within this three-member complex. Here, HDX-MS was used to study the dynamic change of the full-length protein FtsQ, FtsB, and FtsL. Direct experimental evidence shows that FtsB and FtsL form a stable complex by the transmembrane and periplasmic regions. After the complex formation, the structural feature in FtsB properly becomes more defined. This transition may also bring the constriction control domains (CCDs) close to the one in FtsL. Furthermore, with the helical interaction between two proteins, the CCDs have a more determined location relative to the membrane surface. On the other hand, the FtsQ in the FtsQBL complex showed rigidity enhancement in two regions. The interaction with FtsB would fix a significant portion of the FtsQ β-domains. Interestingly, the formation of the FtsQBL complex also stiffens the sequence connecting the two domains in FtsQ. Overall, this study provides important experimental evidence on the conformational dynamic change of the FtsQ, FtsB, and FtsL upon the complex formation and insights into the FtsQBL function in the divisome.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Escherichia Coli

SUBMITTER: Wai Po Kong

LAB HEAD: Kwok-yin Wong

PROVIDER: PXD031342 | Pride | 2022-10-14

REPOSITORIES: Pride

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Publications

The structural dynamics of full-length divisome transmembrane proteins FtsQ, FtsB, and FtsL in FtsQBL complex formation.

Kong Wai-Po WP Gong Furong F So Pui-Kin PK Chen Yu Wai YW Chan Pak-Ho PH Leung Yun-Chung YC Wong Kwok-Yin KY

The Journal of biological chemistry 20220704 8

FtsQBL is a transmembrane protein complex in the divisome of Escherichia coli that plays a critical role in regulating cell division. Although extensive efforts have been made to investigate the interactions between the three involved proteins, FtsQ, FtsB, and FtsL, the detailed interaction mechanism is still poorly understood. In this study, we used hydrogen-deuterium exchange mass spectrometry to investigate these full-length proteins and their complexes. We also dissected the structural dynam ...[more]

PMID: 35798142

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Project description:Acrodysostosis represents a group of rare genetic disorders characterized by defective skeletal development and is often accompanied by intellectual disabilities. Mutations in the 3’5’cyclic AMP (cAMP)-dependent Protein Kinase (PKA) type I regulatory subunit isoform α (RIα) and phosphodiesterase (PDE) PDE4D have both been implicated in impaired PKA regulation in Acrodysostosis. How mutations on PDEs and RIα interfere with regulation of cAMP-PKA signaling is not understood. cAMP-PKA signaling can be described in two phases. In the activation phase, cAMP binding to RIα dissociates the free C-subunit. PDEs hydrolyze cAMP bound to RIα, priming the cAMP-free RIα for reassociation with the C-subunit, thereby completing one PKA activation cycle. Signal termination is thus critical for resetting PKA to its basal state and promoting adaptation to hormonal hyperstimulation. This proceeds through formation of a transient signal termination RIα:PDE complex that facilitates cAMP channeling from the cAMP-binding domain of RIα to the catalytic site of PDE. Signal termination of cAMP-PKA proceeds in three steps: Step 1) Channeling: Translocation of cAMP from the CNB of RIα to the PDE catalytic site for hydrolysis Step 2) Processivity: Binding of free cAMP from the cytosol at both CNBs of RIα and Step 3) Product (5’AMP) release from the PDE hydrolysis site through competitive displacement by a new molecule of cAMP that trigger subsequent activation cycles of PKA. We have identified the molecular basis for two acrodysostosis mutants, (PDE8 T690P) and RIα (T207A) that both allosterically impair cAMP-PKA signal termination. A combination of amide hydrogen/deuterium exchange mass spectrometry (HDXMS) and Fluorescence Polarization (FP) reveal that PDE8 T690P and RIα T207A both blocked processive hydrolysis of cAMP by interfering with competitive displacement of product 5’AMP release from the nucleotide channel at the end of each round of cAMP hydrolysis. While T690P blocked product 5’AMP release from the PDE, T207A greatly slowed release of substrate from RIα. These results highlight the role of processivity in cAMP hydrolysis by RIα:PDE termination complexes for adaptation to cAMP from GPCR hyperstimulation. Impairment of the signal termination process provides an alternate molecular basis for acrodysostosis.

Dataset Information

The structural dynamics of full length divisome transmembrane proteins FtsQ, FtsB, and FtsL in FtsQBL complex formation

Publications

The structural dynamics of full-length divisome transmembrane proteins FtsQ, FtsB, and FtsL in FtsQBL complex formation.

Similar Datasets

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