title: Interactions of histatin-3 and histatin-5 with actin.
abstract: Histatins are histidine rich polypeptides produced in the parotid and submandibular gland and secreted into the saliva. Histatin-3 and -5 are the most important polycationic histatins. They possess antimicrobial activity against fungi such as Candida albicans. Histatin-5 has a higher antifungal activity than histatin-3 while histatin-3 is mostly involved in wound healing in the oral cavity. We found that these histatins, like other polycationic peptides and proteins, such as LL-37, lysozyme and histones, interact with extracellular actin.
  
  
  
  Histatin-3 and -5 polymerize globular actin (G-actin) to filamentous actin (F-actin) and bundle F-actin filaments. Both actin polymerization and bundling by histatins is pH sensitive due to the high histidine content of histatins. In spite of the equal number of net positive charges and histidine residues in histatin-3 and -5, less histatin-3 is needed than histatin-5 for polymerization and bundling of actin. The efficiency of actin polymerization and bundling by histatins greatly increases with decreasing pH. Histatin-3 and -5 induced actin bundles are dissociated by 100 and 50 mM NaCl, respectively. The relatively low NaCl concentration required to dissociate histatin-induced bundles implies that the actin-histatin filaments bind to each other mainly by electrostatic forces. The binding of histatin-3 to F-actin is stronger than that of histatin-5 showing that hydrophobic forces have also some role in histatin-3- actin interaction. Histatins affect the fluorescence of probes attached to the D-loop of G-actin indicating histatin induced changes in actin structure. Transglutaminase cross-links histatins to actin. Competition and limited proteolysis experiments indicate that the main histatin cross-linking site on actin is glutamine-49 on the D-loop of actin.
  
  
  
  Both histatin-3 and -5 interacts with actin, however, histatin 3 binds stronger to actin and affects actin structure at lower concentration than histatin-5 due to the extra 8 amino acid sequence at the C-terminus of histatin-3. Extracellular actin might regulate histatin activity in the oral cavity, which should be the subject of further investigation.
authors.affiliation: Department of Medical Neurobiology, Institute for Medical Research-Israel-Canada, Hebrew University of Jerusalem, Jerusalem, Israel.
authors.forename: Edna
authors.initials: E
authors.lastname: Blotnick
authors.affiliation: Institute of Dental Sciences, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
authors.forename: Asaf
authors.initials: A
authors.lastname: Sol
authors.affiliation: Institute of Dental Sciences, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
authors.forename: Gilad
authors.initials: G
authors.lastname: Bachrach
authors.affiliation: Institute of Dental Sciences, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel. andras.muhlrad@mail.huji.ac.il.
authors.forename: Andras
authors.identifier: 0000-0002-4352-1207
authors.initials: A
authors.lastname: Muhlrad
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000067493:Dynamic Light Scattering; D005456:Fluorescent Dyes / Q000737:chemistry; D054808:Histatins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009994:Osmolar Concentration; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
chemical_list: D000199:Actins; D005456:Fluorescent Dyes; C068075:HTN3 protein, human; D054808:Histatins
keywords: Actin polymerization and bundling; F-actin; G-actin; Histatin-3; Histatin-5; Transglutaminase cross-linking