title: Isolation, biochemical characterization and anti-bacterial activity of BPIFA2 protein.
abstract: OBJECTIVE
  Human BPIFA2 (parotid secretory protein) is a ubiquitous soluble salivary protein, which belongs to the PLUNC family of proteins. Having sequence similarity to bactericidal/permeability-increasing protein and lipopolysaccharide-binding protein, PLUNC proteins are probably involved in local antibacterial response at mucosal sites, such as oral cavity. The aim of the study was to isolate and characterize human BPIFA2.
  
  
  METHODS
  In this paper, we report one-step affinity chromatography method for BPIFA2 purification from whole human saliva. The isolated BPIFA2 was identified by trypsin mass fingerprinting and characterized by electrophoretic methods. Antibacterial activity of BPIFA2 against model microorganism Pseudomonas aeruginosa was shown in minimum inhibitory concentration and time kill study assays.
  
  
  RESULTS
  The protein showed microheterogeneity, both in molecular weight and pI value. BPIFA2 inhibited the growth of P. aeruginosa in microgram concentration range determined by minimum inhibitory concentration assay. In the time kill study, 32μg/mL BPIFA2 showed clear bactericidal activity and did not cause any aggregation of bacteria.
  
  
  CONCLUSIONS
  Affinity chromatography is well suited for isolation of functional BPIFA2 with a potent bactericidal activity against P. aeruginosa.
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Prokopovic
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Milica
authors.initials: M
authors.lastname: Popovic
authors.affiliation: Institute for Chemistry, Technology and Metallurgy, Department of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Uros
authors.initials: U
authors.lastname: Andjelkovic
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Marsavelski
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Brankica
authors.initials: B
authors.lastname: Raskovic
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Gavrovic-Jankulovic
authors.affiliation: Faculty of Chemistry, Department of Biochemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia. Electronic address: polovicn@chem.bg.ac.rs.
authors.forename: Natalija
authors.initials: N
authors.lastname: Polovic
mesh_terms: D000328:Adult; D000372:Agglutination Tests; D002846:Chromatography, Affinity; D005260:Female; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008970:Molecular Weight; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D012463:Saliva / Q000737:chemistry; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: C045623:BPIFA2 protein, human; D012471:Salivary Proteins and Peptides
keywords: BPIFA2; Bactericidal activity; Parotid secretory protein; Saliva