Morphological and proteomic analysis of early stage air-liquid interface biofilm formation in Mycobacterium smegmatis
| Autor: | Zuzana Sochorová, Silvia Bezoušková, Oldřich Benada, Jaroslav Weiser, Karolína Buriánková, Denisa Petráčková, Petr Halada, Barbora Sitařová, Jiří Janeček, Olga Kofroňová |
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| Rok vydání: | 2014 |
| Předmět: |
Gel electrophoresis
Proteomics biology Cell division Scanning electron microscope Mycobacterium smegmatis Air Biofilm Gene Expression Regulation Bacterial biology.organism_classification Mass spectrometry Microbiology Models Biological Bacterial cell structure Culture Media Biochemistry Bacterial Proteins Cell Wall Phase (matter) Biofilms Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Microscopy Electron Scanning Electrophoresis Gel Two-Dimensional |
| Zdroj: | Microbiology (Reading, England). 160(Pt 7) |
| ISSN: | 1465-2080 |
| Popis: | We studied the early stages of pellicle formation by Mycobacterium smegmatis on the surface of a liquid medium [air–liquid interface (A–L)]. Using optical and scanning electron microscopy, we showed the formation of a compact biofilm pellicle from micro-colonies over a period of 8–30 h. The cells in the pellicle changed size and cell division pattern during this period. Based on our findings, we created a model of M. smegmatis A–L early pellicle formation showing the coordinate growth of cells in the micro-colonies and in the homogeneous film between them, where the accessibility to oxygen and nutrients is different. A proteomic approach utilizing high-resolution two-dimensional gel electrophoresis, in combination with mass spectrometry-based protein identification, was used to analyse the protein expression profiles of the different morphological stages of the pellicle. The proteins identified formed four expression groups; the most interesting of these groups contained the proteins with highest expression in the biofilm development phase, when the floating micro-colonies containing long and more robust cells associate into flocs and start to form a compact pellicle. The majority of these proteins, including GroEL1, are involved in cell wall synthesis or modification, mostly through the involvement of mycolic acid biosynthesis, and their expression maxima correlated with the changes in cell size and the rigidity of the bacterial cell wall observed by scanning electron microscopy. |
| Databáze: | OpenAIRE |
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