Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily

Monica Rodriguez De La Vega, Rafael G. Sevilla, Antoni Hermoso, Julia Lorenzo, Sebastian Tanco, Amalia Diez, Lloyd D. Fricker, José M. Bautista, Francesc X. Avilés

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Nna1 has some sequence similarity to metallocarboxypeptidases, but the biochemical characterization of Nna1 has not previously been reported. In this work we performed a detailed genomic scan and found >100 Nna1 homologues in bacteria, Protista, and Animalia, including several paralogs in most eukaryotic species. Phylogenetic analysis of the Nna1-like sequences demonstrates a major divergence between Nna1-like peptidases and the previously known metallocarboxypeptidases subfamilies: M14A, M14B, and M14C. Conformational modeling of representative Nna1-like proteins from a variety of species indicates an unusually open active site, a property that might facilitate its action on a wide variety of peptide and protein substrates. To test this, we expressed a recombinant form of one of the Nna1-like peptidases from Caenorhabditis elegans and demonstrated that this protein is a fully functional metallocarboxypeptidase that cleaves a range of C-terminal amino acids from synthetic peptides. The enzymatic activity is activated by ATP/ADP and salt-inactivated, and is preferentially inhibited by Z-Glu-Tyr dipeptide, which is without precedent in metallocarboxypeptidases and resembles tubulin carboxypeptidase functioning; this hypothesis is strongly reinforced by the results depicted in Kalinina et al. published as accompanying paper in this journal (1). Our findings demonstrate that the M14 family of metallocarboxypeptidases is more complex and diverse than expected, and that Nna1-like peptidases are functional variants of such enzymes, representing a novel subfamily (we propose the name M14D) that contributes substantially to such diversity.

Original languageEnglish (US)
Pages (from-to)851-865
Number of pages15
JournalFASEB Journal
Volume21
Issue number3
DOIs
StatePublished - Mar 2007

Fingerprint

metallocarboxypeptidases
Peptide Hydrolases
peptidases
Caenorhabditis elegans Proteins
Peptides
Proteins
proteins
Dipeptides
Adenosine Diphosphate
Names
Catalytic Domain
Bacteria
carboxypeptidases
Salts
Adenosine Triphosphate
Animalia
dipeptides
synthetic peptides
Amino Acids
Caenorhabditis elegans

Keywords

  • CCP
  • Degradome
  • Peptidase classification
  • Tubulin processing
  • Tubulinyl-Tyr carboxypeptidase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Rodriguez De La Vega, M., Sevilla, R. G., Hermoso, A., Lorenzo, J., Tanco, S., Diez, A., ... Avilés, F. X. (2007). Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily. FASEB Journal, 21(3), 851-865. https://doi.org/10.1096/fj.06-7330com

Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily. / Rodriguez De La Vega, Monica; Sevilla, Rafael G.; Hermoso, Antoni; Lorenzo, Julia; Tanco, Sebastian; Diez, Amalia; Fricker, Lloyd D.; Bautista, José M.; Avilés, Francesc X.

In: FASEB Journal, Vol. 21, No. 3, 03.2007, p. 851-865.

Research output: Contribution to journalArticle

Rodriguez De La Vega, M, Sevilla, RG, Hermoso, A, Lorenzo, J, Tanco, S, Diez, A, Fricker, LD, Bautista, JM & Avilés, FX 2007, 'Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily', FASEB Journal, vol. 21, no. 3, pp. 851-865. https://doi.org/10.1096/fj.06-7330com
Rodriguez De La Vega M, Sevilla RG, Hermoso A, Lorenzo J, Tanco S, Diez A et al. Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily. FASEB Journal. 2007 Mar;21(3):851-865. https://doi.org/10.1096/fj.06-7330com
Rodriguez De La Vega, Monica ; Sevilla, Rafael G. ; Hermoso, Antoni ; Lorenzo, Julia ; Tanco, Sebastian ; Diez, Amalia ; Fricker, Lloyd D. ; Bautista, José M. ; Avilés, Francesc X. / Nna1-like proteins are active metallocarboxypeptidases of a new and diverse M14 subfamily. In: FASEB Journal. 2007 ; Vol. 21, No. 3. pp. 851-865.
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