Purification of Type IV Pili and Pilin Subunits.

Batch Cell Culture Techniques / methods Chemical Fractionation / methods Detergents / chemistry Fimbriae Proteins / chemistry Fimbriae, Bacterial / chemistry Hydrophobic and Hydrophilic Interactions Neisseria gonorrhoeae / chemistry
Neisseria gonorrhoeae PilE Pilin Type IV pili

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2019
Historique:
entrez: 24 5 2019
pubmed: 24 5 2019
medline: 29 2 2020
Statut: ppublish

Résumé

The Type IV pili are displayed peritrichously on the surfaces of Neisseria gonorrhoeae cells. Here we present protocols for isolating and purifying Type IV pili and dissociating them into PilE pilin subunits. Pilus filaments are isolated from the bacterial cell surface by mechanical shearing and purified by differential precipitation and centrifugation. PilE subunits are extracted by treating the purified pili with detergent to disrupt the hydrophobic interactions holding them together in the filaments. Purified pili and pilin subunits can be used for structural, biophysical, or biochemical characterization and as antigens for antibody production.

Identifiants

DOI: 10.1007/978-1-4939-9496-0_7 PMID: 31119620
pubmed: 31119620
doi: 10.1007/978-1-4939-9496-0_7
doi:

Substances chimiques

Detergents 0
pilE protein, Neisseria gonorrhoeae 0
Fimbriae Proteins 147680-16-8

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

97-110

Subventions

Organisme : CIHR
ID : MOP125959
Pays : Canada

Auteurs

Lisa Craig (L)

Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada. licraig@sfu.ca.

Tuba Altindal (T)

Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada.

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Classifications MeSH

questionsmedicales.fr › Techniques d'investigation › Techniques in vitro › Techniques de culture › Techniques de culture cellulaire › Techniques de culture cellulaire en batch › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Fractionnement chimique › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Technologie, industrie et agriculture › Produits domestiques › Détergents › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Protéines de la membrane externe bactérienne › Protéines de fimbriae › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Structures bactériennes › Fimbriae bactériens › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Phénomènes chimiques › Interactions hydrophobes et hydrophiles › Purification of Type IV Pili and Pilin Subunits.
questionsmedicales.fr › Bactéries › Proteobacteria › Betaproteobacteria › Neisseriaceae › Neisseria › Neisseria gonorrhoeae › Purification of Type IV Pili and Pilin Subunits.