PXD064530

PXD064530 is an original dataset announced via ProteomeXchange.

Title	Cleavage preference of metalloprotease EA1
Description	The cleavage preference of the thermophilic metalloprotease from Geobacillus sp. EA1 was characterised using mass spectrometry, confirming its similarity to thermolysin in preferring P1’ hydrophobic amino acids. While EA1 showed similar efficiencies for cleavage of short peptides with one of six hydrophobic amino acids at the P1’ position, thermolysin showed marked preference for leucine. A single amino acid difference in the S1’ pocket of these similar enzymes provides a rationale for this difference. A variant of EA1 (F133L) had intermediary preference, suggesting that approximately half the difference in preference is attributable to this single amino acid change. Broader preference and higher efficiency make EA1 a superior candidate for quick digestion of varied protein substrates.
HostingRepository	PRIDE
AnnounceDate	2025-11-25
AnnouncementXML	Submission_2025-11-25_11:42:39.596.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Torsten Kleffmann
SpeciesList	scientific name: Bacillus thermoproteolyticus; NCBI TaxID: NEWT:1427; scientific name: Escherichia coli; NCBI TaxID: NEWT:562; scientific name: Geobacillus sp. EFT1; NCBI TaxID: NEWT:1108144;
ModificationList	iTRAQ8plex-116 reporter+balance reagent acylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2025-06-02 15:30:16	ID requested
⏵ 1	2025-11-25 11:42:40	announced

10.1002/1873-3468.70123;

submitter keyword: Protease cleavage assay,Thermolysin, iTRAQ

Sigurd Wilbanks
contact affiliation	Department of Biochemistry, University of Otago, New Zealand
contact email	sigurd.wilbanks@otago.ac.nz
lab head
Torsten Kleffmann
contact affiliation	Department of Biochemistry, University of Otago
contact email	torsten.kleffmann@otago.ac.nz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD064530
     1. Label: PRIDE project
     2. Name: Cleavage preference of metalloprotease EA1