PXD010569

PXD010569 is an original dataset announced via ProteomeXchange.

Title	Characterization of serine proteases from Canavalia ensiformis leaf
Description	Serine proteases has been vastly reported to be found (and identified) only in seed extracts of Canavalia ensiformis (C. ensiformis), however, our group observed expressive serine protease activities in aqueous extracts from leaves, seeds, roots, and stem, especially in a leaf extract obtained with distillated water (CE-A). Additionally, this extract demonstrated very good stability in high temperatures and in the presence of chemical agents. In order to study the serine protease activity in leaves, CE-A was submitted to benzamidine-sepharose affinity column and an enriched fraction of serine proteases, named CE-ABza was purified 1.65 fold, yielding a total recovery of about 62%. Based upon zymographic analysis in a gelatin-SDS-PAGE-, the CE-ABza fraction presented an enzymatic activity band at approximately 90 kDa molecular mass region under non-reducing conditions and three bands at 17, 32 and also at 90 kDa under reducing condition. This fraction was able to hydrolyze, in a different way, peptidomimetic and protein substrates. The maximal activity was observed at basic pH values (8.0 to 9.5) and temperature about 40°C. These enzymes exhibited important thermal stability and were significantly inhibited by serine protease inhibitors, such as benzamidine and TPCK. Calcium, magnesium, manganese and zinc ions had a negative modulation on the CE-ABza activity. Mass spectrometry-based proteomics approach identified showed homology with 11 plant proteases from different legumes species, indicating their presence or demonstrating that CE-Bza proteases share specific region of the primary structure of theses enzyme, although the C. ensiformis genome and protein databases are yet to be publicly available. These results demonstrated that these molecules found in both the CE-A extract and the CE-ABza fraction presented important properties that could be used as potential pharmacological and biotechnological targets as therapeutic option. Besides, identification and characterization of C. ensiformis proteases contribute to the studies of plant enzymes.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:10:26.775.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD010569
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Rayane Gon;alves
SpeciesList	scientific name: Canavalia ensiformis; NCBI TaxID: 3823;
ModificationList	Propionamide; Oxidation; Carbamidomethyl
Instrument	Q-Tof micro

Revision	Datetime	Status	ChangeLog Entry
0	2018-07-26 01:50:38	ID requested
1	2020-08-28 07:14:32	announced
2	2020-12-21 03:42:33	announced	2020-12-21: Updated publication reference for DOI(s): https://doi.org/10.1007/s13562-020-00578-5.
⏵ 3	2023-11-14 07:10:27	announced	2023-11-14: Updated project metadata.

10.6019/PXD010569;

curator keyword: Biological

submitter keyword: serine proteases,Canavalia ensiformis, purification

Raquel Elisa da Silva López
contact affiliation	Departamento de Produtos Naturais, Laboratório de Bioquímica de Proteases e Inibidores de Proteases Naturais e Sintéticos, Fundação Oswaldo Cruz (Fiocruz),Brasil
contact email	raquel.lopez@far.fiocruz.br
lab head
Rayane Gon;alves
contact affiliation	Fiocruz
contact email	rayanenatashe@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010569
     1. Label: PRIDE project
     2. Name: Characterization of serine proteases from Canavalia ensiformis leaf