Vertebrate lens α-crystallins are modified by O-linked N-acetylglucosamine

Elizabeth P. Roquemore, Anne Dell, Howard R. Morris, Maria Panico, Andrew J. Reason, Luc Alain Savoy, Graeme J. Wistow, J. Samuel Zigler, Betty J. Earles, Gerald W. Hart

Research output: Contribution to journalArticlepeer-review

90 Scopus citations


Crystallins are structural proteins responsible for establishing the remarkable optical properties of the lens. Yet many of these highly conserved proteins are also expressed in nonocular tissues, where they have alternative functions apparently unrelated to their structural role in the lens. Here we report that lens acrystallins, some of which function as heat-shock proteins in other tissues, are modified with O-linked N-acetylglucosamine (O-GlcNAc). An in vitro enzymatic assay that transfers [3H]Gal to terminal GlcNAc moieties labels αA and αB crystallins in lens homogenates from man, rhesus monkey, rat, cow, and rhea (an ostrich-like bird). O-Linkage of the saccharide is demonstrated by sensitivity to base-catalyzed β-elimination and resistance to peptide:N-glycosidase F treatment. Chromatographic analyses of the β-elimination products and fast atom bombardment-mass spectrometry of [3H]Gal-labeled tryptic peptides confirm the saccharide structure. Isoelectric focusing of [3H]Gal-labeled bovine lens proteins reveals the presence of O-GlcNAc on all four α-crystallin subunits, A1, A2, B1, and B2. Electrospray mass spectrometry of bovine α-crystallin demonstrates the presence of a single O-GlcNAc substitution on αA2. Gas-phase protein sequencing and fast atom bombardment-mass spectrometry of the major radiolabeled tryptic peptide from bovine a-crystallin reveal that GlcNAc is attached to the αA subunits at serine 162. This post-translational modification may play an important role in the molecular organization of lens α-crystallin.

Original languageEnglish (US)
Pages (from-to)555-563
Number of pages9
JournalJournal of Biological Chemistry
Issue number1
StatePublished - 1992
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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