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J Biol Chem. 2013 May 17;288(20):13936-49. doi: 10.1074/jbc.M112.445841. Epub 2013 Mar 21.

Identifying natural substrates for dipeptidyl peptidases 8 and 9 using terminal amine isotopic labeling of substrates (TAILS) reveals in vivo roles in cellular homeostasis and energy metabolism.

Author information

1
School of Biological Sciences, Flinders University, Adelaide, South Australia 5001, Australia.

Abstract

Dipeptidyl peptidases (DP) 8 and 9 are homologous, cytoplasmic N-terminal post-proline-cleaving enzymes that are anti-targets for the development of DP4 (DPPIV/CD26) inhibitors for treating type II diabetes. To date, DP8 and DP9 have been implicated in immune responses and cancer biology, but their pathophysiological functions and substrate repertoire remain unknown. This study utilizes terminal amine isotopic labeling of substrates (TAILS), an N-terminal positional proteomic approach, for the discovery of in vivo DP8 and DP9 substrates. In vivo roles for DP8 and DP9 in cellular metabolism and homeostasis were revealed via the identification of more than 29 candidate natural substrates and pathways affected by DP8/DP9 overexpression. Cleavage of 14 substrates was investigated in vitro; 9/14 substrates for both DP8 and DP9 were confirmed by MALDI-TOF MS, including two of high confidence, calreticulin and adenylate kinase 2. Adenylate kinase 2 plays key roles in cellular energy and nucleotide homeostasis. These results demonstrate remarkable in vivo substrate overlap between DP8/DP9, suggesting compensatory roles for these enzymes. This work provides the first global investigation into DP8 and DP9 substrates, providing a number of leads for future investigations into the biological roles and significance of DP8 and DP9 in human health and disease.

KEYWORDS:

Adenylate Kinase; Aminopeptidase; Calreticulin; Cell Metabolism; DP8/DPP8; DP9/DPP9; Dipeptidyl Peptidase; Energy Metabolism; Enzymes; Proteomics

PMID:
23519473
PMCID:
PMC3656252
DOI:
10.1074/jbc.M112.445841
[Indexed for MEDLINE]
Free PMC Article

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