Abstract

ThefunctionofcytochromesP450canbeclassifiedintotwomajordivisions,thosethatoperateinxenobiotic degradationandthosethatplaycriticalrolesinsteroidhormonesynthesis.Duetothecriticalroleof cytochromeP450inhumanhealth,theseenzymesystemshaveoccupiedNIHsupportedinvestigatorsfor manydecades.Thegenerallongtermgoalsoftheresearchprogramaretogainamolecularlevelinsightinto theimpressiverangeoffunctionaldiversitydisplayedbytheseenzymes.Theessentialfocusofthework proposeddealswithtwoimportanthumanenzymesinvolvedinsteroidbiosynthesis,CYP17andCYP51,both currentdrugtargetsforvariousdiseases,includingatherosclerosisandprostatecancer,progressinthis endeavorultimatelydependsongainingadeeperunderstandingoftheirstructureandprecisereaction pathways.Inordertofullyunderstandthepertinentrelationshipofstructuretofunction,itisimportantto acquirestructuralinformationfornotonlythestableterminalstatesinareaction,butalsoforthereaction intermediates,investigationofthelatterhavingbeenhistoricallyimpeded,owingtotheirfleetingexistence. Also,thefactthatthesearemembrane?boundproteinshascomplicatedtheirinvestigationowingtoproblems associatedwithaggregation.Inordertoaddresstheseobstacles,thisresearchprogramemploysanapproach usinginnovativenanodisctechnologywhichnotonlyprovidesanenvironmentmimickingthenaturalcell membranefortheisolatedCYPenzyme,butalsoenablesthesynthesisofuniquefunctionaldyadsoftheCYP withitsnaturaloralternativereductases.Alsoofimmenseimportance,apowerfulcombinationofresonance Ramanspectroscopywiththecryoradiolysistechniquepermitstrappinganddetailedstructural characterizationofotherwiseelusive,butcrucialreactionintermediates,therebyprovidingaheretofore unattainablelevelofinsightintothestructureandfunctionoftheseenzymes.

Public Health Relevance

ThecytochromesP450playcentralrolesinhumanhealth,oneoftheirfunctionsbeingthebiosynthesisof steroidhormones.Theessentialgoalofthisprojectistogainabetterunderstandingofthestructureand functionoftwohumancytochromeP450drugtargets,CYP17A1andCYP51A1.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM125303-02
Application #
9534143
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Barski, Oleg
Project Start
2017-08-01
Project End
2021-06-30
Budget Start
2018-07-01
Budget End
2019-06-30
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost

  Institution

Name
Marquette University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
046929621
City
Milwaukee
State
WI
Country
United States
Zip Code
53201

  Publications

Mak, Piotr J; Duggal, Ruchia; Denisov, Ilia G et al. (2018) Human Cytochrome CYP17A1: The Structural Basis for Compromised Lyase Activity with 17-Hydroxyprogesterone. J Am Chem Soc 140:7324-7331