Heterogeneity in the M. tuberculosis β-Lactamase Inhibition by Sulbactam.
Marius SchmidtTek Narsingh MallaKara ZielinskiLuis AldamaSasa BajtDenisse FelizBrandon HayesMark S HunterChristopher KupitzStella LisovaJuraj KnoskaJose Manuel Martin-GarciaValerio MarianiSuraj PandeyIshwor PoudyalRaymond G SierraAlexandra TolstikovaOleksandr YefanovChing Hong YoonAbbas OurmazdPetra FrommePeter SchwanderAnton BartyHenry N ChapmanEmina A StojkovicAlexander BatyukSébastien BoutetGeorge N PhilipsLois PollackPublished in: Research square (2023)
For decades, researchers have been determined to elucidate essential enzymatic functions on the atomic lengths scale by tracing atomic positions in real time. Our work builds on new possibilities unleashed by mix-and-inject serial crystallography (MISC) 1-5 at X-ray free electron laser facilities. In this approach, enzymatic reactions are triggered by mixing substrate or ligand solutions with enzyme microcrystals 6 . Here, we report in atomic detail and with millisecond time-resolution how the Mycobacterium tuberculosis enzyme BlaC is inhibited by sulbactam (SUB). Our results reveal ligand binding heterogeneity, ligand gating 7-9 , cooperativity, induced fit 10,11 and conformational selection 11-13 all from the same set of MISC data, detailing how SUB approaches the catalytic clefts and binds to the enzyme non-covalently before reacting to a trans- enamine. This was made possible in part by the application of the singular value decomposition 14 to the MISC data using a newly developed program that remains functional even if unit cell parameters change during the reaction.
Keyphrases
- single cell
- mycobacterium tuberculosis
- electron microscopy
- electronic health record
- acinetobacter baumannii
- hydrogen peroxide
- escherichia coli
- single molecule
- pulmonary tuberculosis
- big data
- high resolution
- high glucose
- multidrug resistant
- quality improvement
- molecular dynamics simulations
- molecular dynamics
- cell therapy
- genome wide
- stem cells
- emergency department
- magnetic resonance imaging
- diabetic rats
- hiv aids
- magnetic resonance
- high speed
- drug resistant
- dna methylation
- drug induced
- bone marrow
- hepatitis c virus
- antiretroviral therapy
- artificial intelligence
- contrast enhanced