Structure and function of glycosylated tandem repeats from Candida albicans Als adhesins

Eukaryot Cell. 2010 Mar;9(3):405-14. doi: 10.1128/EC.00235-09. Epub 2009 Oct 9.

Abstract

Tandem repeat (TR) regions are common in yeast adhesins, but their structures are unknown, and their activities are poorly understood. TR regions in Candida albicans Als proteins are conserved glycosylated 36-residue sequences with cell-cell aggregation activity (J. M. Rauceo, R. De Armond, H. Otoo, P. C. Kahn, S. A. Klotz, N. K. Gaur, and P. N. Lipke, Eukaryot. Cell 5:1664-1673, 2006). Ab initio modeling with either Rosetta or LINUS generated consistent structures of three-stranded antiparallel beta-sheet domains, whereas randomly shuffled sequences with the same composition generated various structures with consistently higher energies. O- and N-glycosylation patterns showed that each TR domain had exposed hydrophobic surfaces surrounded by glycosylation sites. These structures are consistent with domain dimensions and stability measurements by atomic force microscopy (D. Alsteen, V. Dupres, S. A. Klotz, N. K. Gaur, P. N. Lipke, and Y. F. Dufrene, ACS Nano 3:1677-1682, 2009) and with circular dichroism determination of secondary structure and thermal stability. Functional assays showed that the hydrophobic surfaces of TR domains supported binding to polystyrene surfaces and other TR domains, leading to nonsaturable homophilic binding. The domain structures are like "classic" subunit interaction surfaces and can explain previously observed patterns of promiscuous interactions between TR domains in any Als proteins or between TR domains and surfaces of other proteins. Together, the modeling techniques and the supporting data lead to an approach that relates structure and function in many kinds of repeat domains in fungal adhesins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence / genetics
  • Candida albicans / chemistry*
  • Candida albicans / genetics
  • Cell Adhesion Molecules / chemistry*
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism
  • Circular Dichroism
  • Disaccharides / chemistry
  • Enzyme-Linked Immunosorbent Assay
  • Fibronectins / metabolism
  • Fungal Proteins / chemistry*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Glycosylation
  • Lectins / chemistry*
  • Mannosides / chemistry
  • Models, Molecular
  • Molecular Dynamics Simulation
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Polystyrenes / metabolism
  • Protein Binding / physiology
  • Protein Denaturation
  • Protein Interaction Domains and Motifs / physiology*
  • Protein Renaturation
  • Protein Structure, Secondary / physiology
  • Protein Structure, Tertiary
  • Sequence Homology, Amino Acid
  • Serine / chemistry
  • Threonine / chemistry

Substances

  • ALA1 protein, Candida albicans
  • Cell Adhesion Molecules
  • Disaccharides
  • Fibronectins
  • Fungal Proteins
  • Lectins
  • Mannosides
  • Peptide Fragments
  • Polystyrenes
  • mannosyl alpha(1-6)-mannoside
  • Threonine
  • Serine