Structural Insight Into Conformational Changes Induced by ATP Binding in a Type III Secretion-Associated ATPase From Shigella flexneri

Frontiers in Microbiology
Xiaopan GaoSheng Cui

Abstract

Gram-negative bacteria utilize the type III secretion system (T3SS) to inject effector proteins into the host cell cytoplasm, where they subvert cellular functions and assist pathogen invasion. The conserved type III-associated ATPase is critical for the separation of chaperones from effector proteins, the unfolding of effector proteins and translocating them through the narrow channel of the secretion apparatus. However, how ATP hydrolysis is coupled to the mechanical work of the enzyme remains elusive. Herein, we present a complete description of nucleoside triphosphate binding by surface presentation antigens 47 (Spa47) from Shigella flexneri, based on crystal structures containing ATPγS, a catalytic magnesium ion and an ordered water molecule. Combining the crystal structures of Spa47-ATPγS and unliganded Spa47, we propose conformational changes in Spa47 associated with ATP binding, the binding of ATP induces a conformational change of a highly conserved luminal loop, facilitating ATP hydrolysis by the Spa47 ATPase. Additionally, we identified a specific hydrogen bond critical for ATP recognition and demonstrated that, while ATPγS is an ideal analog for probing ATP binding, AMPPNP is a poor ATP mimic. Our findings provide s...Continue Reading

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Citations

Aug 9, 2019·Protein Science : a Publication of the Protein Society·Ivonne BernalMichael Kolbe
Feb 9, 2019·Nature Communications·Dorothy D MajewskiNatalie C J Strynadka
Dec 10, 2020·Microorganisms·Waad BajunaidFrançois-Xavier Campbell-Valois
May 18, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Nicholas J MatzkeMatthew A B Baker
Sep 21, 2021·Frontiers in Molecular Biosciences·Xiaopan GaoSheng Cui

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Methods Mentioned

BETA
electron microscopy
PCR
size exclusion chromatography
isothermal titration calorimetry
size-exclusion chromatography
X-ray
affinity purification
pull-down

Software Mentioned

XDS
PHENIX
Coot
AUTOSHARP
PyMOL
SHARP
Schrödinger
GraphPad Prism
GraphPad
PDBePISA

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