title: Heparin-binding motifs and biofilm formation by Candida albicans.
abstract: Candida albicans is a leading pathogen in infections of central venous catheters, which are frequently infused with heparin. Binding of C. albicans to medically relevant concentrations of soluble and plate-bound heparin was demonstrable by confocal microscopy and enzyme-linked immunosorbent assay (ELISA). A sequence-based search identified 34 C. albicans surface proteins containing ≥1 match to linear heparin-binding motifs. The virulence factor Int1 contained the most putative heparin-binding motifs (n = 5); peptides encompassing 2 of 5 motifs bound to heparin-Sepharose. Alanine substitution of lysine residues K805/K806 in 804QKKHQIHK811 (motif 1 of Int1) markedly attenuated biofilm formation in central venous catheters in rats, whereas alanine substitution of K1595/R1596 in 1593FKKRFFKL1600 (motif 4 of Int1) did not impair biofilm formation. Affinity-purified immunoglobulin G (IgG) recognizing motif 1 abolished biofilm formation in central venous catheters; preimmune IgG had no effect. After heparin treatment of C. albicans, soluble peptides from multiple C. albicans surface proteins were detected, such as Eno1, Pgk1, Tdh3, and Ssa1/2 but not Int1, suggesting that heparin changes candidal surface structures and may modify some antigens critical for immune recognition. These studies define a new mechanism of biofilm formation for C. albicans and a novel strategy for inhibiting catheter-associated biofilms.
authors.affiliation: Division of Pediatric Infectious Diseases.
authors.forename: Julianne V
authors.initials: JV
authors.lastname: Green
authors.forename: Kris I
authors.initials: KI
authors.lastname: Orsborn
authors.forename: Minlu
authors.initials: M
authors.lastname: Zhang
authors.forename: Queenie K G
authors.initials: QK
authors.lastname: Tan
authors.forename: Kenneth D
authors.initials: KD
authors.lastname: Greis
authors.forename: Alexey
authors.initials: A
authors.lastname: Porollo
authors.forename: David R
authors.initials: DR
authors.lastname: Andes
authors.forename: Jason
authors.initials: J
authors.lastname: Long Lu
authors.forename: Margaret K
authors.initials: MK
authors.lastname: Hostetter
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D018441:Biofilms; D002176:Candida albicans / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D062905:Central Venous Catheters / Q000382:microbiology; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D051381:Rats
chemical_list: D005656:Fungal Proteins; D008565:Membrane Proteins; D006493:Heparin
references.citation: AIDS. 1994 Feb;8(2):183-92
references.pmid: 8043225
references.citation: PLoS Pathog. 2006 Jul;2(7):e63
references.pmid: 16839200
references.citation: Yeast. 2004 Mar;21(4):285-302
references.pmid: 15042589
references.citation: J Biol Chem. 1993 Sep 15;268(26):19228-31
references.pmid: 8366075
references.citation: Gene. 1987;57(2-3):267-72
references.pmid: 3319781
references.citation: Antimicrob Agents Chemother. 2012 Jan;56(1):148-53
references.pmid: 21986822
references.citation: J Biol Chem. 1992 May 5;267(13):8857-62
references.pmid: 1577724
references.citation: Cell Microbiol. 2006 Sep;8(9):1382-91
references.pmid: 16848788
references.citation: J Biol Chem. 2004 Dec 31;279(53):55744-53
references.pmid: 15501822
references.citation: Eukaryot Cell. 2008 Nov;7(11):1906-15
references.pmid: 18723603
references.citation: J Bacteriol. 1992 Nov;174(21):6789-99
references.pmid: 1400228
references.citation: Proteomics. 2004 Oct;4(10):3084-106
references.pmid: 15378761
references.citation: Bioinformatics. 2005 May 1;21(9):1846-52
references.pmid: 15691858
references.citation: Anal Biochem. 2002 Nov 15;310(2):199-207
references.pmid: 12423639
references.citation: Arteriosclerosis. 1989 Jan-Feb;9(1):21-32
references.pmid: 2463827
references.citation: Glycobiology. 2009 Dec;19(12):1537-46
references.pmid: 19729381
references.citation: Microbiol Mol Biol Rev. 2008 Sep;72(3):495-544
references.pmid: 18772287
references.citation: Clin Pharm. 1988 Jun;7(6):439-43
references.pmid: 3402178
references.citation: J Mol Biol. 2004 Jul 16;340(4):783-95
references.pmid: 15223320
references.citation: J Infect Dis. 2008 Apr 1;197(7):981-9
references.pmid: 18419534
references.citation: Cell. 2012 Jan 20;148(1-2):126-38
references.pmid: 22265407
references.citation: Chem Biol. 2005 Mar;12(3):267-77
references.pmid: 15797210
references.citation: Virulence. 2013 Feb 15;4(2):119-28
references.pmid: 23302789
references.citation: Am J Physiol Gastrointest Liver Physiol. 2009 Feb;296(2):G266-74
references.pmid: 19033532
references.citation: PLoS Pathog. 2010 Mar 26;6(3):e1000827
references.pmid: 20361054
references.citation: J Infect Dis. 1993 May;167(5):1247-51
references.pmid: 8486965
references.citation: BMC Bioinformatics. 2008 Sep 23;9:392
references.pmid: 18811934
references.citation: J Biol Chem. 1999 Oct 1;274(40):28198-205
references.pmid: 10497173
references.citation: PLoS One. 2012;7(8):e42692
references.pmid: 22880084
references.citation: Science. 1998 Feb 27;279(5355):1355-8
references.pmid: 9478896
references.citation: Clin Infect Dis. 2009 Jul 1;49(1):1-45
references.pmid: 19489710
references.citation: Proteomics. 2001 Apr;1(4):550-9
references.pmid: 11681208
references.citation: Intensive Care Med. 2004 Apr;30(4):633-8
references.pmid: 14722639
references.citation: J Biol Chem. 2003 Aug 1;278(31):28553-61
references.pmid: 12761219
references.citation: Infect Immun. 2004 Oct;72(10):6023-31
references.pmid: 15385506
references.citation: J Biol Chem. 2011 Jul 1;286(26):23608-19
references.pmid: 21572042
references.citation: Angew Chem Int Ed Engl. 2002 Feb 1;41(3):391-412
references.pmid: 12491369
references.citation: Nephron Clin Pract. 2007;107(4):c128-32
references.pmid: 17957123
references.citation: Infect Immun. 2005 Aug;73(8):4596-606
references.pmid: 16040971
references.citation: Cell Mol Biol. 1991;37(3):253-60
references.pmid: 1934005
references.citation: J Bacteriol. 1999 Mar;181(6):1868-74
references.pmid: 10074081
references.citation: Eukaryot Cell. 2007 Apr;6(4):693-709
references.pmid: 17220463