title: Re-evaluation of a bacterial antifreeze protein as an adhesin with ice-binding activity.
abstract: A novel role for antifreeze proteins (AFPs) may reside in an exceptionally large 1.5-MDa adhesin isolated from an Antarctic Gram-negative bacterium, Marinomonas primoryensis. MpAFP was purified from bacterial lysates by ice adsorption and gel electrophoresis. We have previously reported that two highly repetitive sequences, region II (RII) and region IV (RIV), divide MpAFP into five distinct regions, all of which require mM Ca(2+) levels for correct folding. Also, the antifreeze activity is confined to the 322-residue RIV, which forms a Ca(2+)-bound beta-helix containing thirteen Repeats-In-Toxin (RTX)-like repeats. RII accounts for approximately 90% of the mass of MpAFP and is made up of ∼120 tandem 104-residue repeats. Because these repeats are identical in DNA sequence, their number was estimated here by pulsed-field gel electrophoresis. Structural homology analysis by the Protein Homology/analogY Recognition Engine (Phyre2) server indicates that the 104-residue RII repeat adopts an immunoglobulin beta-sandwich fold that is typical of many secreted adhesion proteins. Additional RTX-like repeats in RV may serve as a non-cleavable signal sequence for the type I secretion pathway. Immunodetection shows both repeated regions are uniformly distributed over the cell surface. We suggest that the development of an AFP-like domain within this adhesin attached to the bacterial outer surface serves to transiently bind the host bacteria to ice. This association would keep the bacteria within the upper reaches of the water column where oxygen and nutrients are potentially more abundant. This novel envirotactic role would give AFPs a third function, after freeze avoidance and freeze tolerance: that of transiently binding an organism to ice.
authors.affiliation: Protein Function Discovery Group and the Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada.
authors.forename: Shuaiqi
authors.initials: S
authors.lastname: Guo
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Garnham
authors.forename: John C
authors.initials: JC
authors.lastname: Whitney
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Graham
authors.forename: Peter L
authors.initials: PL
authors.lastname: Davies
mesh_terms: D018829:Adhesins, Bacterial / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D000327:Adsorption; D000595:Amino Acid Sequence; D021301:Antifreeze Proteins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D015139:Blotting, Southern; D005455:Fluorescent Antibody Technique; D015698:Genomic Library; D007053:Ice; D053544:Marinomonas / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D053719:Tandem Mass Spectrometry; D020080:Tandem Repeat Sequences
chemical_list: D018829:Adhesins, Bacterial; D021301:Antifreeze Proteins; D007053:Ice