Imperial College London
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Dr Josefin Ahnström

Faculty of Medicine, Department of Immunology and Inflammation

Senior Lecturer in Haematology
 
 
 
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Contact

 

+44 (0)20 3313 4236j.ahnstrom

 
 
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Location

 

5S5Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Martin:2021:10.1016/j.jprot.2021.104358,
author = {Martin, DR and Santamaria, S and Koch, CD and Ahnström, J and Apte, SS},
doi = {10.1016/j.jprot.2021.104358},
journal = {Journal of Proteomics},
pages = {1--8},
title = {Identification of novel ADAMTS1, ADAMTS4 and ADAMTS5 cleavage sites in versican using a label-free quantitative proteomics approach.},
url = {http://dx.doi.org/10.1016/j.jprot.2021.104358},
volume = {249},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - The chondroitin sulfate proteoglycan versican is important for embryonic development and several human disorders. The versican V1 splice isoform is widely expressed and cleaved by ADAMTS proteases at a well-characterized site, Glu441-Ala442. Since ADAMTS proteases cleave the homologous proteoglycan aggrecan at multiple sites, we hypothesized that additional cleavage sites existed within versican. We report a quantitative label-free approach that ranks abundance of liquid chromatography-tandem mass spectrometry (LC-MS/MS)-identified semi-tryptic peptides after versican digestion by ADAMTS1, ADAMTS4 and ADAMTS5 to identify site-specific cleavages. Recombinant purified versican V1 constructs were digested with the recombinant full-length proteases, using catalytically inactive mutant proteases in control digests. Semi-tryptic peptide abundance ratios determined by LC-MS/MS in ADAMTS:control digests were compared to the mean of all identified peptides to obtain a z-score by which outlier peptides were ranked, using semi-tryptic peptides identifying Glu441 -Ala442 cleavage as the benchmark. Tryptic peptides with higher abundance in control:ADAMTS digests supported cleavage site identification. We identified several novel cleavage sites supporting the ADAMTS1/4/5 cleavage site preference for a P1-Glu residue in proteoglycan substrates. Digestion of proteins in vitro and application of this z-score approach is potentially widely applicable for mapping protease cleavage sites using label-free proteomics. SIGNIFICANCE: Versican abundance and turnover are relevant to the pathogenesis of several human disorders. Versican is cleaved by A Disintegrin-like And Metalloprotease with Thrombospondin type 1 motifs (ADAMTS) family members at Glu441-Ala442, generating a bioactive proteoform called versikine, but additional cleavage sites and the site-specificity of individual ADAMTS proteases is unexplored. Here, we used label-free proteomics approach to identify versican cleavage sites
AU  - Martin,DR
AU  - Santamaria,S
AU  - Koch,CD
AU  - Ahnström,J
AU  - Apte,SS
DO  - 10.1016/j.jprot.2021.104358
EP  - 8
PY  - 2021///
SN  - 0165-022X
SP  - 1
TI  - Identification of novel ADAMTS1, ADAMTS4 and ADAMTS5 cleavage sites in versican using a label-free quantitative proteomics approach.
T2  - Journal of Proteomics
UR  - http://dx.doi.org/10.1016/j.jprot.2021.104358
UR  - https://www.ncbi.nlm.nih.gov/pubmed/34450332
UR  - https://www.sciencedirect.com/science/article/pii/S1874391921002578?via%3Dihub
UR  - http://hdl.handle.net/10044/1/91843
VL  - 249
ER  -
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