Immunoelectron microscopic studies on the localization of peptidoglycan peptide subunit pentapeptides in bacterial cell walls |
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Authors: | Peter H. Seidl Jochen R. Golecki Norbert Franken Karl Heinz Schleifer |
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Affiliation: | (1) Lehrstuhl für Mikrobiologie, Technische Universität München, Arcisstraß 21, D-8000 München 2, Federal Republic of Germany;(2) Institut für Biologie II, Mikrobiologie, Albert-Ludwigs-Universität, Schänzlestraße 1, D-7800 Freiburg i. Br., Federal Republic of Germany |
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Abstract: | The peptide subunit pentapeptide H-L-Ala-D-Glu(L-Lys-D-Ala-D-Ala-OH)-NH2 of peptidoglycan was localized in the cell walls of several Gram-positive bacteria employing the indirect immunoferritin technique. Specific antibodies to the D-alanyl-D-alanine moiety of non-crosslinked peptide subunit pentapeptide were raised in rabbits by immunization with synthetic immunogen albumin-(CH2CO-Gly-L-Ala-L-Ala-D-Ala-D-Ala-OH)39. Specificity of these antibodies for the peptide subunit pentapeptide and not for the peptide subunit tetrapeptide was corroborated in a Farr-type radio-active hapten binding assay. Specificity of labelling with ferritin was established by immunoelectron microscopic controls, and by the excellent correlation between specific labelling of cells with ferritin and the particular peptidoglycan primary structure of bacterial strains investigated. Cells of Lactobacillus gasseri, Streptococcus pyogenes and Staphylococcus aureus revealing non-crosslinked peptide subunit pentapeptides in their peptidoglycans could specifically be labelled. Lactobacillus acidophilus and Bacillus subtilis, on the contrary, missing such pentapeptides, failed in labelling.The implication of this method to possibly localize the points of attack of penicillin or cycloserine is discussed.Abbreviations used meso-A2pm meso-diaminopimelic acid - DSM Deutsche Sammlung für Mikroorganismen, Göttingen, FRGThis paper is dedicated to Professor Gerhart Drews on the occasion of his 60th birthday |
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Keywords: | Indirect immunoferritin technique Peptidoglycan Synthetic antigens |
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