Chapter title |
Analysis of Bacterial Pilus Assembly by Shearing and Immunofluorescence Microscopy
|
---|---|
Chapter number | 18 |
Book title |
Protein Complex Assembly
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7759-8_18 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7758-1, 978-1-4939-7759-8
|
Authors |
Areli Luna-Rico, Jenny-Lee Thomassin, Olivera Francetic, Luna-Rico, Areli, Thomassin, Jenny-Lee, Francetic, Olivera |
Abstract |
Bacterial surface appendages of the type 4 pilus superfamily play diverse roles in adherence, aggregation, motility, signaling, and macromolecular transport. Here we describe two analytical approaches to study assembly of type 4 pili and of pseudopili produced by type 2 protein secretion systems: the shearing assay and immunofluorescence microscopy. These complementary antibody-based methods allow for semiquantitative analysis of fiber assembly. The shearing assay can be scaled up to yield crude extracts of pili that can be further analyzed by electron and atomic force microscopy or by mass spectrometry. |
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