title: Conformational preferences of linear beta-defensins are revealed by ion mobility-mass spectrometry.
abstract: In recent times there has been an enormous rise in resistance to synthetic antibiotics as well as an increase in the virulence of bacteria, the so-called "superbugs". This problem has catalyzed a search for novel molecules to fight bacteria, which in turn relies on a better understanding of the molecular basis of the immune response. Beta-defensins are a class of small, cationic, cysteine-rich antimicrobial peptides expressed by humans and other animals to act against incoming pathogens. As well as their antimicrobial properties, beta-defensins also act as chemokines, recruiting cells to the sites of infection. Here the relationship between the tertiary structures of beta-defensin analogs and their chemotactic activities has been investigated using ion mobility-mass spectrometry (IM-MS) and biochemical assays. A panel of derivatives of the murine beta-defensin Defb14 has been formed and the ability of these peptides to chemoattract the receptor CCR6 has been assessed in vitro. The derivatives can be divided into two groups, those with chemotactic activity equal to that of the unmodified parent peptide, and those whose chemotactic activity has been lost upon modification. Analysis by ion mobility-mass spectrometry reveals the conformational preferences of these peptides upon ionization from different solvents. Under denaturing conditions, the chemotactic peptides adopt more compact conformations in the gas-phase at higher charge states than those which are inactive. While the conditions of these experiments are not akin to the environment around the receptor in vivo, this technique provides an in vacuo method for distinguishing between the different chemotactic activities of beta-defensin derivatives.
authors.affiliation: School of Chemistry, University of Edinburgh, Edinburgh, EH9 3JJ, UK.
authors.forename: Martin
authors.initials: M
authors.lastname: De Cecco
authors.forename: Emily S
authors.initials: ES
authors.lastname: Seo
authors.forename: David J
authors.initials: DJ
authors.lastname: Clarke
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: McCullough
authors.forename: Karen
authors.initials: K
authors.lastname: Taylor
authors.forename: Derek
authors.initials: D
authors.lastname: Macmillan
authors.forename: Julia R
authors.initials: JR
authors.lastname: Dorin
authors.forename: Dominic J
authors.initials: DJ
authors.lastname: Campopiano
authors.forename: Perdita E
authors.initials: PE
authors.lastname: Barran
mesh_terms: D002460:Cell Line; D006801:Humans; D007477:Ions / Q000737:chemistry*; D013058:Mass Spectrometry; D011487:Protein Conformation; D011489:Protein Denaturation; D054399:Receptors, CCR6 / Q000378:metabolism; D023083:beta-Defensins / Q000737:chemistry*
chemical_list: C106311:CCR6 protein, human; D007477:Ions; D054399:Receptors, CCR6; D023083:beta-Defensins