Newborn mouse lens proteome and its alteration by lysine 6 mutant ubiquitin

Fu Shang, Phillip Wilmarth, Min Lee Chang, Ke Liu, Larry David, Maria Andrea Caceres, Eric Wawrousek, Allen Taylor

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Ubiquitin is a tag that often initiates degradation of proteins by the proteasome in the ubiquitin proteasome system. Targeted expression of K6W mutant ubiquitin (K6W-Ub) in the lens results in defects in lens development and cataract formation, suggesting critical functions for ubiquitin in lens. To study the developmental processes that require intact ubiquitin, we executed the most extensive characterization of the lens proteome to date. We quantified lens protein expression changes in multiple replicate pools of P1 wild-type and K6W-Ub-expressing mouse lenses. Lens proteins were digested with trypsin, peptides were separated using strong cation exchange and reversed-phase liquid chromatography, and tandem mass (MS/MS) spectra were collected with a linear ion trap. Transgenic mice that expressed low levels of K6W-Ub (low expressers) had normal, clear lenses at birth, whereas the lenses that expressed high levels of K6W-Ub (higher expressers) had abnormal lenses and cataracts at birth. A total of 2052 proteins were identified, of which 996 were reliably quantified and compared between wild-type and K6W-Ub transgenic mice. Consistent with a delayed developmental program, fiber-cell-specific proteins, such as γ-crystallins (γA, γB, γC, and γE), were down-regulated in K6W-Ub higher expressers. Up-regulated proteins were involved in energy metabolism, signal transduction, and proteolysis. The K6W-Ub low expressers exhibited delayed onset and milder cataract consistent with smaller changes in protein expression. Because lens protein expression changes occurred prior to lens morphological abnormalities and cataract formation in K6W-Ub low expressers, it appears that expression of K6W-Ub sets in motion a process of altered protein expression that results in developmental defects and cataract.

Original languageEnglish (US)
Pages (from-to)1177-1189
Number of pages13
JournalJournal of Proteome Research
Volume13
Issue number3
DOIs
StatePublished - Mar 7 2014

Fingerprint

Proteome
Ubiquitin
Lenses
Lysine
Crystallins
Cataract
Proteins
Proteasome Endopeptidase Complex
Transgenic Mice
Proteolysis
Parturition
Signal transduction
Defects
Liquid chromatography
Reverse-Phase Chromatography
Trypsin
Energy Metabolism
Cations
Signal Transduction
Ions

Keywords

  • mass spectrometry
  • mouse lens
  • nuclear cataract
  • proteasome
  • proteomics
  • ubiquitin

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Newborn mouse lens proteome and its alteration by lysine 6 mutant ubiquitin. / Shang, Fu; Wilmarth, Phillip; Chang, Min Lee; Liu, Ke; David, Larry; Caceres, Maria Andrea; Wawrousek, Eric; Taylor, Allen.

In: Journal of Proteome Research, Vol. 13, No. 3, 07.03.2014, p. 1177-1189.

Research output: Contribution to journalArticle

Shang, F, Wilmarth, P, Chang, ML, Liu, K, David, L, Caceres, MA, Wawrousek, E & Taylor, A 2014, 'Newborn mouse lens proteome and its alteration by lysine 6 mutant ubiquitin', Journal of Proteome Research, vol. 13, no. 3, pp. 1177-1189. https://doi.org/10.1021/pr400801v
Shang, Fu ; Wilmarth, Phillip ; Chang, Min Lee ; Liu, Ke ; David, Larry ; Caceres, Maria Andrea ; Wawrousek, Eric ; Taylor, Allen. / Newborn mouse lens proteome and its alteration by lysine 6 mutant ubiquitin. In: Journal of Proteome Research. 2014 ; Vol. 13, No. 3. pp. 1177-1189.
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AU - Wawrousek, Eric

AU - Taylor, Allen

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AB - Ubiquitin is a tag that often initiates degradation of proteins by the proteasome in the ubiquitin proteasome system. Targeted expression of K6W mutant ubiquitin (K6W-Ub) in the lens results in defects in lens development and cataract formation, suggesting critical functions for ubiquitin in lens. To study the developmental processes that require intact ubiquitin, we executed the most extensive characterization of the lens proteome to date. We quantified lens protein expression changes in multiple replicate pools of P1 wild-type and K6W-Ub-expressing mouse lenses. Lens proteins were digested with trypsin, peptides were separated using strong cation exchange and reversed-phase liquid chromatography, and tandem mass (MS/MS) spectra were collected with a linear ion trap. Transgenic mice that expressed low levels of K6W-Ub (low expressers) had normal, clear lenses at birth, whereas the lenses that expressed high levels of K6W-Ub (higher expressers) had abnormal lenses and cataracts at birth. A total of 2052 proteins were identified, of which 996 were reliably quantified and compared between wild-type and K6W-Ub transgenic mice. Consistent with a delayed developmental program, fiber-cell-specific proteins, such as γ-crystallins (γA, γB, γC, and γE), were down-regulated in K6W-Ub higher expressers. Up-regulated proteins were involved in energy metabolism, signal transduction, and proteolysis. The K6W-Ub low expressers exhibited delayed onset and milder cataract consistent with smaller changes in protein expression. Because lens protein expression changes occurred prior to lens morphological abnormalities and cataract formation in K6W-Ub low expressers, it appears that expression of K6W-Ub sets in motion a process of altered protein expression that results in developmental defects and cataract.

KW - mass spectrometry

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KW - nuclear cataract

KW - proteasome

KW - proteomics

KW - ubiquitin

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