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PXD053893

PXD053893 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleDistinct mechanistic, conformational and substrate selectivity profiles emerging in the evolution of high catalytic activity enzymes via parallel trajectories
DescriptionRecent laboratory evolution studies have demonstrated that parallel evolutionary trajectories can lead to genetically distinct enzymes with high activity towards a non-preferred substrate. However, it is unknown whether such genetically distinct enzymes have convergent conformational dynamics and mechanistic features. To address this question, we used as a model the Homo sapiens kynureninase (HsKYNase), which is of great interest for cancer immunotherapy. Earlier, we reported on a highly unusual evolutionary trajectory that led to the isolation of HsKYNase_66, having a 410-fold increase in the kcat/KM for kynurenine (KYN) and reverse substrate selectivity relative to the parental wild-type enzyme. Here, by following a completely different evolutionary trajectory we generated a genetically distinct variant, HsKYNase_93D9, that exhibits KYN catalytic activity comparable to that of HsKYNase_66, but instead it is a “generalist” that accepts 3’-hydroxykynurenine (OH-KYN) with the same proficiency. Pre-steady-state kinetic analysis revealed that whereas the evolution of HsKYNase_66 was accompanied by a change in the rate-determining step of the reaction, HsKYNase_93D9 retained the same catalytic mechanism as the parental OH-KYN-preferring human enzyme. HDX-MS revealed that the conformational dynamics of the two enzymes during catalysis are markedly different and distinct from ortholog procaryotic enzymes with high KYN activity. Our work provides a mechanistic framework for understanding the relationship betweenevolutionary mechanisms and phenotypic traits of evolved generalist and specialist enzyme species.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_06:51:42.709.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterSheena D'Arcy
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentSynapt MS
Dataset History
RevisionDatetimeStatusChangeLog Entry
02024-07-11 15:51:49ID requested
12024-07-29 08:07:03announced
22024-10-22 06:51:43announced2024-10-22: Updated project metadata.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: kynureninase, enzyme, substrate,HDX, engineering
Contact List
Sheena D'Arcy
contact affiliationDepartment of Chemistry and Biochemistry, The University of Texas at Dallas
contact emailsheena.darcy@utdallas.edu
lab head
Sheena D'Arcy
contact affiliationThe University of Texas at Dallas
contact emailsheena.darcy@utdallas.edu
dataset submitter
Full Dataset Link List
Dataset FTP location
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