In Vivo and In Vitro Protein-Peptidoglycan Interactions

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1615; p. 143
• Main Authors: Li, Gang, Peter Howard, S
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Electrophoresis, Polyacrylamide Gel; Gram-Negative Bacteria; Membrane Proteins; Peptidoglycan - chemistry; Peptidoglycan - isolation & purification; Peptidoglycan - metabolism; Protein Binding; Muramic acid assay; Peptidoglycan; Trans-envelope systems; Co-sedimentation; Cross-linking
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-7033-9_11

Bacteria have developed a number of trans-envelope systems to transport molecules or assemble organelles across bacterial envelopes. However, bacterial envelopes contain a rigid netlike peptidoglycan structure that protects cells from osmotic lysis. Trans-envelope systems thus must interact with the peptidoglycan barrier to generate gaps or anchor structures to the peptidoglycan scaffold. Here we describe methods to use in vivo cross-linking and in vitro co-sedimentation to study protein-peptidoglycan interactions in Gram-negative bacteria. In particular, we address important considerations to ensure the specificity of the interactions in question.