Zymographic Characterization of Staphylococcus aureus Cell Wall

• Published in: MICROBIOLOGY and IMMUNOLOGY 1998, Vol.42(3), pp.231-235
• Main Authors: Ohta, Kouji, Komatsuzawa, Hitoshi, Sugai, Motoyuki, Suginaka, Hidekazu
• Format: Article
• Language: English
• Subjects: Amidohydrolases - metabolism; Anti-Bacterial Agents - pharmacology; Autolysin; Bacteriological methods and techniques used in bacteriology; Bacteriology; Bacteriolysis; Biological and medical sciences; Cell wall; Cell Wall - chemistry; Cell Wall - drug effects; Cell Wall - metabolism; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Hexosaminidases - metabolism; Lysostaphin - metabolism; MBD; Microbiology; Muramidase - metabolism; Peptidoglycan - chemistry; Peptidoglycan - drug effects; Peptidoglycan - metabolism; Staphylococcus aureus - chemistry; Staphylococcus aureus - drug effects; Zymography
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1998.tb02276.x

Susceptibilities of several preparations of Staphylococcus aureus cells to various peptidoglycan hydrolases with known bond specificity were analyzed by zymography. The substrates were intact S. aureus cells, cells boiled in the presence of SDS and cells treated with trichloroacetic acid after treatment with boiling SDS solution (TCA-cells). Twofold dilution of lysostaphin (LS), lysozyme (LZ), S. aureus 51kDa glucosaminidase (GL) or S. aureus 62kDa amidase (AM) were electrophoresed, and the minimal enzyme dose showing a visible bacteriolytic band was defined as MBD (minimal bacteriolytic dose). Under the same experimental conditions, this method gave reproducible results. As the substrate for zymogram, TCA-cells were the most sensitive to LS, LZ and AM, whereas the three substrate were equally sensitive to GL. A zymographic analysis of methicillin-resistant S. aureus treated with methicillin together with previous studies suggest that this method can be used for the preliminary characterization of S. aureus cell wall peptidoglycan.