Cytosolic carboxypeptidase 1 is involved in processing α- and β-tubulin

Iryna Berezniuk, Hang T. Vu, Peter J. Lyons, Juan J. Sironi, Hui Xiao, Berta Burd, Mitsutoshi Setou, Ruth H. Angeletti, Koji Ikegami, Lloyd D. Fricker

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The Purkinje cell degeneration (pcd) mouse has a disruption in the gene encoding cytosolic carboxypeptidase 1 (CCP1). This study tested two proposed functions of CCP1: degradation of intracellular peptides and processing of tubulin. Overexpression (2-3-fold) or knockdown (80-90%) of CCP1 in human embryonic kidney 293T cells (HEK293T) did not affect the levels of most intracellular peptides but altered the levels of α-tubulin lacking two C-terminal amino acids (delta2-tubulin) ≥5-fold, suggesting that tubulin processing is the primary function of CCP1, not peptide degradation. Purified CCP1 produced delta2-tubulin from purified porcine brain α-tubulin or polymerized HEK293T microtubules. In addition, CCP1 removed Glu residues from the polyglutamyl side chains of porcine brain α- and β-tubulin and also generated a form of α-tubulin with two C-terminal Glu residues removed (delta3-tubulin). Consistent with this, pcd mouse brain showed hyperglutamylation of both α- and β-tubulin. The hyperglutamylation of α- and β-tubulin and subsequent death of Purkinje cells in pcd mice was counteracted by the knock-out of the gene encoding tubulin tyrosine ligase-like-1, indicating that this enzyme hyperglutamylates α- and β-tubulin. Taken together, these results demonstrate a role for CCP1 in the processing of Glu residues from β- as well as α-tubulin in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)6503-6517
Number of pages15
JournalJournal of Biological Chemistry
Volume287
Issue number9
DOIs
StatePublished - Feb 24 2012

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Carboxypeptidases
Tubulin
Processing
Purkinje Cells
Brain
Gene encoding
HEK293 Cells
Peptides
Swine
Kidney
Degradation
Gene Knockout Techniques
Microtubules

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Cytosolic carboxypeptidase 1 is involved in processing α- and β-tubulin. / Berezniuk, Iryna; Vu, Hang T.; Lyons, Peter J.; Sironi, Juan J.; Xiao, Hui; Burd, Berta; Setou, Mitsutoshi; Angeletti, Ruth H.; Ikegami, Koji; Fricker, Lloyd D.

In: Journal of Biological Chemistry, Vol. 287, No. 9, 24.02.2012, p. 6503-6517.

Research output: Contribution to journalArticle

Berezniuk, I, Vu, HT, Lyons, PJ, Sironi, JJ, Xiao, H, Burd, B, Setou, M, Angeletti, RH, Ikegami, K & Fricker, LD 2012, 'Cytosolic carboxypeptidase 1 is involved in processing α- and β-tubulin', Journal of Biological Chemistry, vol. 287, no. 9, pp. 6503-6517. https://doi.org/10.1074/jbc.M111.309138
Berezniuk I, Vu HT, Lyons PJ, Sironi JJ, Xiao H, Burd B et al. Cytosolic carboxypeptidase 1 is involved in processing α- and β-tubulin. Journal of Biological Chemistry. 2012 Feb 24;287(9):6503-6517. https://doi.org/10.1074/jbc.M111.309138
Berezniuk, Iryna ; Vu, Hang T. ; Lyons, Peter J. ; Sironi, Juan J. ; Xiao, Hui ; Burd, Berta ; Setou, Mitsutoshi ; Angeletti, Ruth H. ; Ikegami, Koji ; Fricker, Lloyd D. / Cytosolic carboxypeptidase 1 is involved in processing α- and β-tubulin. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 9. pp. 6503-6517.
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