The role of the SEA (sea urchin sperm protein, enterokinase and agrin) module in cleavage of membrane-tethered mucins

Timea Palmai-Pallag, Naila Khodabukus, Leo Kinarsky, Shih Hsing Leir, Simon Sherman, Michael A Hollingsworth, Ann Harris

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The membrane-tethered mucins are cell surface-associated dimeric or multimeric molecules with extracellular, transmembrane and cytoplasmic portions, that arise from cleavage of the primary polypeptide chain. Following the first cleavage, which may be cotranslational, the subunits remain closely associated through undefined noncovalent interactions. These mucins all share a common structural motif, the SEA module that is found in many other membrane-associated proteins that are released from the cell surface and has been implicated in both the cleavage events and association of the subunits. Here we examine the SEA modules of three membrane-tethered mucins, MUC1, MUC3 and MUC12, which have significant sequence homology within the SEA domain. We previously identified the primary, cleavage site within the MUC1 SEA domain as FRPG/SVVV a sequence that is highly conserved in MUC3 and MUC12. We now show by site-directed mutagenesis that the F, G and S residues are important for the efficiency of the cleavage reaction but not indispensable and that amino acids outside this motif are probably important. These data are consistent with a new model of the MUC1 SEA domain that is based on the solution structure of the MUC16 SEA module, derived by NMR spectroscopy. Further, we demonstrate that cleavage of human MUC3 and MUC12 occurs within the SEA domain. However, the SEA domains of MUC1, MUC3 and MUC12 are not interchangeable, suggesting that either these modules alone are insufficient to mediate efficient cleavage or that the 3D structure of the hybrid molecules does not adequately recreate an accessible cleavage site.

Original languageEnglish (US)
Pages (from-to)2901-2911
Number of pages11
JournalFEBS Journal
Volume272
Issue number11
DOIs
StatePublished - Jun 1 2005

Fingerprint

Agrin
Enteropeptidase
Sea Urchins
Mucins
Spermatozoa
Membranes
Mutagenesis
Amino Acid Motifs
Proteins
Molecules
Sequence Homology
Site-Directed Mutagenesis
Nuclear magnetic resonance spectroscopy
Membrane Proteins
Magnetic Resonance Spectroscopy
Amino Acids
Peptides

Keywords

  • MUC12
  • MUC3
  • Proteolytic cleavage
  • SEA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The role of the SEA (sea urchin sperm protein, enterokinase and agrin) module in cleavage of membrane-tethered mucins. / Palmai-Pallag, Timea; Khodabukus, Naila; Kinarsky, Leo; Leir, Shih Hsing; Sherman, Simon; Hollingsworth, Michael A; Harris, Ann.

In: FEBS Journal, Vol. 272, No. 11, 01.06.2005, p. 2901-2911.

Research output: Contribution to journalArticle

Palmai-Pallag, Timea ; Khodabukus, Naila ; Kinarsky, Leo ; Leir, Shih Hsing ; Sherman, Simon ; Hollingsworth, Michael A ; Harris, Ann. / The role of the SEA (sea urchin sperm protein, enterokinase and agrin) module in cleavage of membrane-tethered mucins. In: FEBS Journal. 2005 ; Vol. 272, No. 11. pp. 2901-2911.
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