title: Novel entries in a fungal biofilm matrix encyclopedia.
abstract: Virulence of Candida is linked with its ability to form biofilms. Once established, biofilm infections are nearly impossible to eradicate. Biofilm cells live immersed in a self-produced matrix, a blend of extracellular biopolymers, many of which are uncharacterized. In this study, we provide a comprehensive analysis of the matrix manufactured by Candida albicans both in vitro and in a clinical niche animal model. We further explore the function of matrix components, including the impact on drug resistance. We uncovered components from each of the macromolecular classes (55% protein, 25% carbohydrate, 15% lipid, and 5% nucleic acid) in the C. albicans biofilm matrix. Three individual polysaccharides were identified and were suggested to interact physically. Surprisingly, a previously identified polysaccharide of functional importance, β-1,3-glucan, comprised only a small portion of the total matrix carbohydrate. Newly described, more abundant polysaccharides included α-1,2 branched α-1,6-mannans (87%) associated with unbranched β-1,6-glucans (13%) in an apparent mannan-glucan complex (MGCx). Functional matrix proteomic analysis revealed 458 distinct activities. The matrix lipids consisted of neutral glycerolipids (89.1%), polar glycerolipids (10.4%), and sphingolipids (0.5%). Examination of matrix nucleic acid identified DNA, primarily noncoding sequences. Several of the in vitro matrix components, including proteins and each of the polysaccharides, were also present in the matrix of a clinically relevant in vivo biofilm. Nuclear magnetic resonance (NMR) analysis demonstrated interaction of aggregate matrix with the antifungal fluconazole, consistent with a role in drug impedance and contribution of multiple matrix components. Importance: This report is the first to decipher the complex and unique macromolecular composition of the Candida biofilm matrix, demonstrate the clinical relevance of matrix components, and show that multiple matrix components are needed for protection from antifungal drugs. The availability of these biochemical analyses provides a unique resource for further functional investigation of the biofilm matrix, a defining trait of this lifestyle.
authors.affiliation: Department of Medicine, Infectious Diseases, University of Wisconsin, Madison, Wisconsin, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Zarnowski
authors.affiliation: National Magnetic Resonance Facility at Madison, University of Wisconsin, Madison, Wisconsin, USA.
authors.forename: William M
authors.initials: WM
authors.lastname: Westler
authors.affiliation: Department of Biochemistry, University of Wisconsin, Madison, Wisconsin, USA.
authors.forename: Ghislain Ade
authors.initials: GA
authors.lastname: Lacmbouh
authors.affiliation: Dairy Forage Research Center, U.S. Department of Agriculture, Madison, Wisconsin, USA.
authors.forename: Jane M
authors.initials: JM
authors.lastname: Marita
authors.affiliation: Department of Biochemistry, University of Wisconsin, Madison, Wisconsin, USA.
authors.forename: Jameson R
authors.initials: JR
authors.lastname: Bothe
authors.forename: Jörg
authors.initials: J
authors.lastname: Bernhardt
authors.affiliation: Université du Littoral Côte d'Opale, Unité de Chimie Environnementale at Interactions sur le Vivant, Calais, France.
authors.forename: Anissa
authors.initials: A
authors.lastname: Lounes-Hadj Sahraoui
authors.affiliation: Université du Littoral Côte d'Opale, Unité de Chimie Environnementale at Interactions sur le Vivant, Calais, France.
authors.forename: Joël
authors.initials: J
authors.lastname: Fontaine
authors.affiliation: Department of Medicine, Infectious Diseases, University of Wisconsin, Madison, Wisconsin, USA.
authors.forename: Hiram
authors.initials: H
authors.lastname: Sanchez
authors.affiliation: Dairy Forage Research Center, U.S. Department of Agriculture, Madison, Wisconsin, USA.
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Hatfield
authors.forename: James M
authors.initials: JM
authors.lastname: Ntambi
authors.forename: Jeniel E
authors.initials: JE
authors.lastname: Nett
authors.affiliation: Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Mitchell
authors.affiliation: dra@medicine.wisc.edu.
authors.forename: David R
authors.initials: DR
authors.lastname: Andes
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D002176:Candida albicans; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D040901:Proteomics / Q000379:methods*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins
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