title: Escherichia coli expression, purification, crystallization, and structure determination of bacterial cohesin-dockerin complexes.
abstract: Cellulosomes are highly efficient nanomachines that play a fundamental role during the anaerobic deconstruction of complex plant cell wall carbohydrates. The assembly of these complex nanomachines results from the very tight binding of repetitive cohesin modules, located in a noncatalytic molecular scaffold, and dockerin domains located at the C-terminus of the enzyme components of the cellulosome. The number of enzymes found in a cellulosome varies but may reach more than 100 catalytic subunits if cellulosomes are further organized in polycellulosomes, through a second type of cohesin-dockerin interaction. Structural studies have revealed how the cohesin-dockerin interaction mediates cellulosome assembly and cell-surface attachment, while retaining the flexibility required to potentiate catalytic synergy within the complex. Methods that might be applied for the production, purification, and structure determination of cohesin-dockerin complexes are described here.
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Pólo Universitário do Alto da Ajuda, Lisboa, Portugal.
authors.forename: Joana L A
authors.initials: JL
authors.lastname: Brás
authors.forename: Ana Luisa
authors.initials: AL
authors.lastname: Carvalho
authors.forename: Aldino
authors.initials: A
authors.lastname: Viegas
authors.forename: Shabir
authors.initials: S
authors.lastname: Najmudin
authors.forename: Victor D
authors.initials: VD
authors.lastname: Alves
authors.forename: José A M
authors.initials: JA
authors.lastname: Prates
authors.forename: Luís M A
authors.initials: LM
authors.lastname: Ferreira
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Romão
authors.forename: Harry J
authors.initials: HJ
authors.lastname: Gilbert
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
mesh_terms: D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D044603:Cellulosomes / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular / Q000187:drug effects*; D048013:Acetivibrio thermocellus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017434:Protein Structure, Tertiary; D000097722:Cohesins
chemical_list: D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D009097:Multienzyme Complexes; D000097722:Cohesins