PXD030867

PXD030867 is an original dataset announced via ProteomeXchange.

Title	Cytochrome P450s protect against oxidative damage via a preferred orientation of the heme cofactor
Description	Cytochrome P450 enzymes (P450s) are a superfamily of monooxygenases that utilise a cysteine thiolate ligated heme moiety to perform a wide range of demanding oxidative transformations. Given the oxidative power of active intermediate formed within P450s during their active cycle, it is remarkable that these enzymes can avoid self-oxidation as well as retaining the axial cysteine ligand in the deprotonated – and thus highly acidic – thiolate form. Whilst little is known about the process of heme incorporation during P450 folding, there is an overwhelming preference for one heme orientation within the P450 active site. Indeed, very few structures to date contain an alternate heme orientation, of which two are OxyA homologues from glycopeptide antibiotic (GPA) biosynthesis. Given the apparent preference for the unusual heme orientation shown by OxyA enzymes, we investigated the OxyA homologue from kistamicin biosynthesis, which is an atypical GPA. We determined that OxyAkis is highly sensitive to oxidative damage by peroxide, with both UV an EPR measurements shows a rapid bleaching of the heme signal. We determined the structure of OxyAkis and found a mixed population of heme orientations present in this enzyme. Analysis further revealed that an unprecedented oxidative modification of the heme was detected, which that correlated with the presence of the alternate heme orientation in the protein. These results provide evidence that the typical heme orientation in Cytochrome P450s can help to prevent potential autocatalytic modification of the heme – and hence deactivation of the enzyme – during P450 catalysis. It also suggests that some P450 enzymes involved in GPA biosynthesis may be especially prone to oxidative damage due to the heme orientation found in their active sites.
HostingRepository	PRIDE
AnnounceDate	2022-05-20
AnnouncementXML	Submission_2022-05-20_04:54:51.207.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ralf Schittenhelm
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-10 04:06:42	ID requested
⏵ 1	2022-05-20 04:54:52	announced

Greule A, Izor, é T, Machell D, Hansen MH, Schoppet M, De Voss JJ, Charkoudian LK, Schittenhelm RB, Harmer JR, Cryle MJ, The Cytochrome P450 OxyA from the Kistamicin Biosynthesis Cyclization Cascade is Highly Sensitive to Oxidative Damage. Front Chem, 10():868240(2022) [pubmed]

submitter keyword: Cytochrome P450s, glycopeptide antibiotic (GPA) biosynthesis

Max Cryle
contact affiliation	Infection and Immunity Program, Monash Biomedicine Discovery Institute, Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, 3800 Australia
contact email	max.cryle@monash.edu
lab head
Ralf Schittenhelm
contact affiliation	Monash University
contact email	ralf.schittenhelm@monash.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD030867
     1. Label: PRIDE project
     2. Name: Cytochrome P450s protect against oxidative damage via a preferred orientation of the heme cofactor