PXD039681-2

PXD039681 is an original dataset announced via ProteomeXchange.

Title	Peptide and MUC5B hydrolysis by the Mucin Degrading Protease (MdpL) from Limosilactobacillus fermentum
Description	MUC5B is the predominant glycoprotein in saliva and is instrumental in the establishment and maintenance of multi-species eubiotic biofilms in the oral cavity. Investigations of the aciduric Lactobacillaceae family, and its role in biofilms emphasizes the diversity across different genera of the proteolytic systems involved in the nutritional utilization of mucins. We have characterized a cysteine protease from Limosilactobacillus fermentum, MdpL (Mucin degrading protease from Limosilactobacillus) with a high protein backbone similarity with commensals that exploit mucins for attachment and nutrition. MdpL was shown to be associated with the bacterial cell surface, in close proximity to MUC5B, which was sequentially degraded into low molecular weight fragments. Mapping the substrate preference revealed multiple hydrolytic sites of proteins with a high O-glycan occurrence, although hydrolysis was not dependent on the presence of O-glycans. However, since proteolysis of immunoglobulins was absent, and general protease activity was low, a preference for glycoproteins similar to MUC5B in terms of glycosylation and structure is suggested. MdpL preferentially hydrolyzed C-terminally located hydrophobic residues in peptides larger than 20 amino acids, which hinted at a limited sequence preference. To secure proper enzyme folding and optimal conditions for activity, L. fermentum incorporates a complex system that establishes a reducing environment. The importance of overall reducing conditions was confirmed by the activity boosting effect of the added reducing agents L-cysteine and DTT. High activity was retained in low to neutral pH 5.5-7.0, but the enzyme was completely inhibited in the presence of Zn2+. Here we have characterized a highly conserved mucin degrading protease from L. fermentum. MdpL, that together with the recently discovered O-glycanase and O-glycoprotease enzyme groups, increases our understanding of mucin degradation and complex biofilm dynamics.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:00:21.295.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Fredrik Leo
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	impact II; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-01-26 02:09:19	ID requested
1	2023-03-22 01:12:00	announced
⏵ 2	2023-11-14 09:00:21	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: IOB, MdpL, Limosilactobacillus fermentum, MUC5B, Neuromedin, MOG,LC-MSMS

Claes Wickström
contact affiliation	Department of Oral Biology and Pathology, Faculty of Odontology, Malmö University, Malmö, Sweden
contact email	claes.wickstrom@mau.se
lab head
Fredrik Leo
contact affiliation	Malmö University
contact email	fredrik.leo@mau.se
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/03/PXD039681

PRIDE project URI

• PRIDE
  1. PXD039681
     1. Label: PRIDE project
     2. Name: Peptide and MUC5B hydrolysis by the Mucin Degrading Protease (MdpL) from Limosilactobacillus fermentum