Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis

Hui Zhang, Sadiqa Maqsudi, Adam Rainczuk, Nadine Duffield, Josie Michelle Lawrence, Fiona M Keane, Daniela Justa-Schuch, Ruth Geiss-Friedlander, Mark Douglas Gorrell, Andrew Stephens

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Dipeptidyl Peptidase 9 (DPP9) is a member of the S9B/DPPIV (DPP4) serine protease family, which cleaves N-terminal dipeptides at an Xaa-Pro consensus motif. Cytoplasmic DPP9 has roles in epidermal growth factor signalling and in antigen processing, whilst the role of the recently discovered nuclear form of DPP9 is unknown. Mice lacking DPP9 proteolytic activity die as neonates. We applied a modified 2D-DIGE approach to identify novel DPP9 substrates, using mouse embryonic fibroblasts lacking endogenous DPP9 activity. A total of 111 potential new DPP9 substrates were identified, with nine proteins/peptides confirmed as DPP9 substrates by MALDI-TOF or immunoblotting. Moreover, we also identified the dipeptide Val-Ala as a consensus site for DPP9 cleavage that was not recognised by DPP8, suggesting different in vivo roles for these closely related enzymes. The relative kinetics for the cleavage of these nine candidate substrates by DPP9, DPP8 and DPP4 were determined. This is the first identification of DPP9 substrates from cells lacking endogenous DPP9 activity. These data greatly expand the potential roles of DPP9 and suggest different in vivo roles for DPP9 and DPP8. This article is protected by copyright. All rights reserved
Original languageEnglish
Pages (from-to)3737 - 3757
Number of pages21
JournalFEBS Journal
Volume282
Issue number19
DOIs
Publication statusPublished - 2015

Cite this

Zhang, Hui ; Maqsudi, Sadiqa ; Rainczuk, Adam ; Duffield, Nadine ; Lawrence, Josie Michelle ; Keane, Fiona M ; Justa-Schuch, Daniela ; Geiss-Friedlander, Ruth ; Gorrell, Mark Douglas ; Stephens, Andrew. / Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis. In: FEBS Journal. 2015 ; Vol. 282, No. 19. pp. 3737 - 3757.
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title = "Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis",
abstract = "Dipeptidyl Peptidase 9 (DPP9) is a member of the S9B/DPPIV (DPP4) serine protease family, which cleaves N-terminal dipeptides at an Xaa-Pro consensus motif. Cytoplasmic DPP9 has roles in epidermal growth factor signalling and in antigen processing, whilst the role of the recently discovered nuclear form of DPP9 is unknown. Mice lacking DPP9 proteolytic activity die as neonates. We applied a modified 2D-DIGE approach to identify novel DPP9 substrates, using mouse embryonic fibroblasts lacking endogenous DPP9 activity. A total of 111 potential new DPP9 substrates were identified, with nine proteins/peptides confirmed as DPP9 substrates by MALDI-TOF or immunoblotting. Moreover, we also identified the dipeptide Val-Ala as a consensus site for DPP9 cleavage that was not recognised by DPP8, suggesting different in vivo roles for these closely related enzymes. The relative kinetics for the cleavage of these nine candidate substrates by DPP9, DPP8 and DPP4 were determined. This is the first identification of DPP9 substrates from cells lacking endogenous DPP9 activity. These data greatly expand the potential roles of DPP9 and suggest different in vivo roles for DPP9 and DPP8. This article is protected by copyright. All rights reserved",
author = "Hui Zhang and Sadiqa Maqsudi and Adam Rainczuk and Nadine Duffield and Lawrence, {Josie Michelle} and Keane, {Fiona M} and Daniela Justa-Schuch and Ruth Geiss-Friedlander and Gorrell, {Mark Douglas} and Andrew Stephens",
year = "2015",
doi = "10.1111/febs.13371",
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Zhang, H, Maqsudi, S, Rainczuk, A, Duffield, N, Lawrence, JM, Keane, FM, Justa-Schuch, D, Geiss-Friedlander, R, Gorrell, MD & Stephens, A 2015, 'Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis' FEBS Journal, vol. 282, no. 19, pp. 3737 - 3757. https://doi.org/10.1111/febs.13371

Identification of novel dipeptidyl peptidase 9 substrates by two-dimensional differential in-gel electrophoresis. / Zhang, Hui; Maqsudi, Sadiqa; Rainczuk, Adam; Duffield, Nadine; Lawrence, Josie Michelle; Keane, Fiona M; Justa-Schuch, Daniela; Geiss-Friedlander, Ruth; Gorrell, Mark Douglas; Stephens, Andrew.

In: FEBS Journal, Vol. 282, No. 19, 2015, p. 3737 - 3757.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Zhang, Hui

AU - Maqsudi, Sadiqa

AU - Rainczuk, Adam

AU - Duffield, Nadine

AU - Lawrence, Josie Michelle

AU - Keane, Fiona M

AU - Justa-Schuch, Daniela

AU - Geiss-Friedlander, Ruth

AU - Gorrell, Mark Douglas

AU - Stephens, Andrew

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AB - Dipeptidyl Peptidase 9 (DPP9) is a member of the S9B/DPPIV (DPP4) serine protease family, which cleaves N-terminal dipeptides at an Xaa-Pro consensus motif. Cytoplasmic DPP9 has roles in epidermal growth factor signalling and in antigen processing, whilst the role of the recently discovered nuclear form of DPP9 is unknown. Mice lacking DPP9 proteolytic activity die as neonates. We applied a modified 2D-DIGE approach to identify novel DPP9 substrates, using mouse embryonic fibroblasts lacking endogenous DPP9 activity. A total of 111 potential new DPP9 substrates were identified, with nine proteins/peptides confirmed as DPP9 substrates by MALDI-TOF or immunoblotting. Moreover, we also identified the dipeptide Val-Ala as a consensus site for DPP9 cleavage that was not recognised by DPP8, suggesting different in vivo roles for these closely related enzymes. The relative kinetics for the cleavage of these nine candidate substrates by DPP9, DPP8 and DPP4 were determined. This is the first identification of DPP9 substrates from cells lacking endogenous DPP9 activity. These data greatly expand the potential roles of DPP9 and suggest different in vivo roles for DPP9 and DPP8. This article is protected by copyright. All rights reserved

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