Pseudomonas aeruginosa

Microbial Expression technology Dr. Horst Funken & Dr. Filip Kovacic │ Institute of Molecular Enzyme Technology │ Heinrich-Heine University Düsseldorf, Forschungszentrum Jülich GmbH │ 52426 Jülich

Pseudomonas aeruginosa

Identification of novel enzymes in P. aeruginosa

 is an aerobic, motile, Gram-negative rod

database search

 Is an opportunistic human pathogen  is associated with chronic

cloning

airway infection

(www.pseudomonas.com)

 is resistant to many antibiotics

homologous & heterologous expression

Proteins we are interested in:  lectins

protein purification

 lipases  esterases activity assays, biochemical characterization, physiological investigation, structure determination,

 phospholipases  proteases

Characterization of virulence factors

Investigation of Pseudomonas Lectins Lectins

Motility assays

 are sugar-binding proteins  function as central mediators of

motility of P. aeruginosa wilde type

information transfer in biological

Biofilm formation

systems

Structure of LecB from P. aeruginosa2

 play an important role in cell-cell

Siderophore production

Pyoverdine detection under UV-light

interactions

Biofilm of P. aeruginosa1

 are secreted by unknown mechanism

Activity assays

PAO1 Localization of LecB by Western Blot3

P. aeruginosa LecB Lipase activity assay

 is located on the cell surface

Proteom investigation

interaction with the outer memrane

2D gel electrophoresis of P. aeruginosa

porin OprF

Microarray analysis

 is an N-glycosylated protein

Hemagglutination of rabbit red blood cells4

 can agglutinate erythrocytes

DNA microarray of P. aeruginosa

References: 1) 2) 3) 4)

Tielker et al., 2005; „Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation” Loris et al., 2003; “Structural basis of carbohydrate recognition by the lectin LecB from Pseudomonas aeruginosa.” Bartels et al., 2011; “Glycosylation Is Required for Outer Membrane Localization of the Lectin LecB in Pseudomonas aeruginosa” Funken et al., 2012; “Specific Association of Lectin LecB with the Surface of Pseudomonas aeruginosa: Role of Outer Membrane Protein OprF”