Peptidase substrates via global peptide profiling

Tagore, Debarati M.; Nolte, Whitney M.; Neveu, John M.; Rangel, Roberto; Guzman-Rojas, Liliana; Pasqualini, Renata; Arap, Wadih; Lane, William S.; Saghatelian, Alan

doi:10.1038/nchembio.126

Cited by 59 publications

(115 citation statements)

References 17 publications

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“…Identification of APN substrates implicated in tumor growth and metastasis is another required next step. A number of cytokines, growth factors, and extracellular matrix molecules implicated in angiogenesis have been reported as putative APN substrates (46)(47)(48)(49)(50)(51)(52)(53)(54)(55), and testing their possible roles in the models presented here will be the focus of future studies.…”

Section: Discussionmentioning

confidence: 99%

Cooperative effects of aminopeptidase N (CD13) expressed by nonmalignant and cancer cells within the tumor microenvironment

Guzman-Rojas

Rangel

Salameh

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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117

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Processes that promote cancer progression such as angiogenesis require a functional interplay between malignant and nonmalignant cells in the tumor microenvironment. The metalloprotease aminopeptidase N (APN; CD13) is often overexpressed in tumor cells and has been implicated in angiogenesis and cancer progression. Our previous studies of APN-null mice revealed impaired neoangiogenesis in model systems without cancer cells and suggested the hypothesis that APN expressed by nonmalignant cells might promote tumor growth. We tested this hypothesis by comparing the effects of APN deficiency in allografted malignant (tumor) and nonmalignant (host) cells on tumor growth and metastasis in APNnull mice. In two independent tumor graft models, APN activity in both the tumors and the host cells cooperate to promote tumor vascularization and growth. Loss of APN expression by the host and/ or the malignant cells also impaired lung metastasis in experimental mouse models. Thus, cooperation in APN expression by both cancer cells and nonmalignant stromal cells within the tumor microenvironment promotes angiogenesis, tumor growth, and metastasis.lung cancer | melanoma | proteolytic activity | shRNA | tumorigenesis A minopeptidase N (APN, CD13; EC 3.4.11.2) is a widely expressed type II membrane-bound metalloprotease (1, 2). It functions in the enzymatic cleavage of peptides, in endocytosis, and as a signaling molecule and has been implicated in the regulation of complex and diverse processes, including cell migration, cell survival, viral uptake, and angiogenesis (3). APN has also been linked specifically to cancer, having been identified as a cellsurface marker for malignant myeloid cells (4-7) and reaching high levels of expression in association with the progression of tumors, including breast, ovarian, and prostate cancer (8-14). Indeed, vascular endothelial growth factor (VEGF), a key angiogenesis regulator, induces the expression of APN at an early stage of tumor growth (15), again highlighting the role of this enzyme in angiogenesis, a process crucial for sustained growth of most solid tumors (16). Studies of bestatin, a CD13 inhibitor and antiangiogenic agent, also suggest that APN enzymatic activity is relevant for tumorigenesis (17,18). Nevertheless, the substrates of APN in the context of angiogenesis are still unknown. The only well-defined substrate is angiotensin III in the renin-angiotensin pathway, in which APN cleaves the NH 2 -terminal arginine residue of angiotensin III to form angiotensin IV. Consistent with several lines of evidence, we have previously identified APN as a target for inhibition of tumor vascularization and growth (18)(19)(20).Tumor growth relies on a complex microenvironment in which malignant cells cooperate with various other cell types: endothelial cells of the blood and lymphatic circulation, mesenchymal stromal cells/cancer-associated fibroblasts, and a variety of bone marrow-derived cells such as myeloid-derived suppressor cells and lymphocytes (21, 22). Some of these cell populations c...

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Section: Discussionmentioning

confidence: 99%

Cooperative effects of aminopeptidase N (CD13) expressed by nonmalignant and cancer cells within the tumor microenvironment

Guzman-Rojas

Rangel

Salameh

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

117

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“…For example, in vitro assays with recombinant enzyme cannot account for protein-protein interactions or spatial localization that can promote or prevent interactions between peptidases and candidate substrates (22). To overcome these challenges, recent studies have begun to look at the direct impact of peptidase activity on the peptides in cells and tissues through the application of mass spectrometry (MS)-based peptidomics (23)(24)(25)(26)(27)(28). In these experiments, changes in the peptide profile as a function of peptidase activity can reveal substrates, products, and pathways regulated by a peptidase in vivo.…”

Section: Introductionmentioning

confidence: 99%

“…In such experiments, known endogenous peptides and, importantly, previously unidentified peptides, including posttranslationally modified versions of known peptides, can be measured (29). Recently, peptidomics has been applied to identify peptidase-regulated peptides, including substrates, by quantifying changes in the tissue peptidome as a function of peptidase activity (23)(24)(25)27,28,31,32) (Fig. 2).…”

Section: Introductionmentioning

confidence: 99%

“…2). Since these initial efforts, similar approaches have been used to study other peptidases as well (23)(24)(25)(26)(27)(28)31,32). Fig.…”

Section: Introductionmentioning

confidence: 99%

“…With Prep, these studies led to an improved understanding of the biochemistry of the enzyme, and illustrated the difficulty in predicting peptidase substrates or cleavage sites based on peptide primary sequence alone (23). In the case of DPP4, peptidomics provided biochemical information about this enzyme and also revealed the existence of interlinked biochemical pathways involved in renal peptide catabolism (25). Lastly, peptide changes regulated by DPP4 were useful as a metric for increasing the coverage of the peptidomics platform through a systematic analysis of the key steps in the workflow (24).…”

Section: Introductionmentioning

confidence: 99%

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Peptidomics of the Prolyl Peptidases

Lone

Nolte

Tinoco

et al. 2010

AAPS J

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Abstract. The prolyl peptidases are a family of enzymes characterized by a biochemical preference for cleaving proline-containing peptides. The members of this enzyme family include prolyl endopeptidase, prolyl endopeptidase-like, dipeptidyl peptidase 4 (DPP4), DPP7, DPP8, DPP9, and fibroblast activation protein. DPP4 is the best studied member of the family, due to its role in physiological glucose tolerance, exerted through the regulation of the insulinotropic peptide glucagon-like peptide-1. While other members of the prolyl peptidase family have also been implicated in various (patho)physiological processes, the underlying peptides and pathways regulated by these enzymes are less clear. The identification of endogenous substrates of the prolyl peptidases is an important step in elucidating the molecular mechanisms of these enzymes. Here, we highlight the utility of liquid chromatography-mass spectrometry-based peptidomics to enable the discovery of endogenous prolyl peptidase substrates directly from tissues, and demonstrate the utility of this information in understanding the biochemical and physiological functions of the prolyl peptidases.

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Naturstoffe und ihre biologischen Angriffsziele: proteomische und metabolomische Markierungsstrategien

2010

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Das aktivitätsbasierte Protein‐Profiling (ABPP) hat sich zu einer ausgereiften Standardmethode für die schnelle, empfindliche und selektive Identifizierung von Enzymaktivitäten und Inhibitoren in Proteomen entwickelt. Mit naturstoffbasierenden Sonden lassen sich die Angriffsziele (targets) vieler bislang uncharakterisierter Moleküle leicht in komplexen Proteomen aufklären und so ihre genaue Funktion und der Wirkmechanismus verstehen. Zum anderen dienen Naturstoffsonden und ihre Derivate als pharmakologische Leitstrukturen, die essenzielle Komponenten in der Zelle hemmen und in biologischen Assays ihre Wirksamkeit zeigen. Da die komplexen regulatorischen Prozesse einer Zelle mehr umfassen als nur Transkription, Translation und Aktivierung, ist es entscheidend, auch die Produkte des aktiven Proteoms – die Metaboliten und Bindungspartner einzelner Enzyme und Proteine – zu identifizieren. Dabei sind Methoden nötig, mit denen sich das chemisch komplexe Metabolom charakterisieren lässt. In den letzten Jahren gab es hierzu eine Reihe interessanter Ansätze, mit denen eine globale Untersuchung von Enzym‐Metabolit‐Paaren jetzt zum ersten Mal möglich ist.

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Peptidase substrates via global peptide profiling

Cited by 59 publications

References 17 publications

Cooperative effects of aminopeptidase N (CD13) expressed by nonmalignant and cancer cells within the tumor microenvironment

Cooperative effects of aminopeptidase N (CD13) expressed by nonmalignant and cancer cells within the tumor microenvironment

Peptidomics of the Prolyl Peptidases

Naturstoffe und ihre biologischen Angriffsziele: proteomische und metabolomische Markierungsstrategien

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