PXD014658

PXD014658 is an original dataset announced via ProteomeXchange.

Title	Mapping of transglutaminase 2 active sites of human salivary small basic proline-rich
Description	Due to the distinctive features of oral mucosa, determination of the proteins involved in the formation of “oral protein pellicle” is demanding. The present study investigated the susceptibility of several human basic proline-rich peptides (bPRPs) named P-H, P-D, P-F, P-J, and II-2 as substrates of transglutaminase-2. The reactivity of P-C peptide and statherin was also investigated. Peptides purified from human whole saliva were incubated at diverse temperatures with two different transglutaminases (type 2; guinea pig and human), in the presence or in the absence of monodansyl-cadaverine. High resolution HPLC-ESI-MS and MS/MS analyses of the reaction products highlighted that P-H, and P-D were active substrates of transglutaminase-2, II-2 was less reactive and the reactivity of P-F and P-J was negligible. All the peptides formed cyclo-derivatives, and only specific glutamine residues were involved in the cycle formation and reacted with monodansyl-cadaverine. Q29 for P-H, Q37 for P-D, and Q21 for II-2 were the principal reactive residues. One or two secondary glutamine residues were hierarchically susceptible to reaction with dansyl-cadaverine. The main consensus sequence recognized by transglutaminase-2 in basic proline-rich peptides was GNPQ, and, despite the great similarity of P-F and P-J with the other salivary peptides, their reactivity was negligible, probably because the GNPQ consensus sequence was missing in their sequence. The reactivity of P-D A32 variant was lower than that of P-D P32 peptide suggesting that the more rigid peptide structure was a better substrates for transglutaminase-2. The reactivity of P-C was high, but followed rules different from the other basic peptides under study, Q41 being the principal reactive residue, confirming that P-C is not strictly related to the basic proline-rich proteins. The reactivity of statherin was very high too, and the manual inspection of the MS/MS fragmentation patterns of the reaction products confirmed, as determined in previous studies, that Q37 is the pivotal residue, Q39 and Q40 the secondary residues recognized by transglutaminase-2. Moreover, Q39 and Q40 residues were more reactive in cyclo-statherin Q37 than in statherin, suggesting that cycle formation increases the reactivity of the secondary glutamine residues. Finally, the physiological temperature (37°C) was the best for the reaction and no significant qualitative differences were observed using human or guinea pig transglutaminase-2.
HostingRepository	PRIDE
AnnounceDate	2019-10-25
AnnouncementXML	Submission_2019-10-25_04:22:44.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Tiziana Cabras
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	N6-(L-isoglutamyl)-L-lysine (Gln)
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2019-07-18 03:14:14	ID requested
1	2019-10-25 00:01:17	announced
⏵ 2	2019-10-25 04:22:45	announced	2019-10-25: Updated publication reference for PubMed record(s): 31638822.

Boroumand M, Olianas A, Manconi B, Serrao S, Iavarone F, Desiderio C, Pieroni L, Faa G, Messana I, Castagnola M, Cabras T, Mapping of Transglutaminase-2 Sites of Human Salivary Small Basic Proline-Rich Proteins by HPLC-High-Resolution ESI-MS/MS. J Proteome Res, 19(1):300-313(2020) [pubmed]

submitter keyword: human, saliva, bPRPs, Transglutaminase 2

Tiziana Cabras
contact affiliation	University of Cagliari, Dep. Life and Environmental Sciences, Biomedical Section
contact email	tcabras@unica.it
lab head
Tiziana Cabras
contact affiliation	University of Cagliari, DISVA
contact email	tcabras@unica.it
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/2019/10/PXD014658

PRIDE project URI

• PRIDE
  1. PXD014658
     1. Label: PRIDE project
     2. Name: Mapping of transglutaminase 2 active sites of human salivary small basic proline-rich