Citation

BibTex format

@article{Matthews:2016:10.1107/S2053230X16017921,
author = {Matthews, SJ and rouse, S and hawthorne and Lambert and hare and morgan, M},
doi = {10.1107/S2053230X16017921},
journal = {Acta Crystallographica Section F: Structural Biology Communications},
pages = {892--896},
title = {Purification, crystallization and characterization of the Pseudomonas outer membrane protein FapF, a functional amyloid transporter},
url = {http://dx.doi.org/10.1107/S2053230X16017921},
volume = {F72},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Bacteria often produce extracellular amyloid fibresviaa multi-componentsecretion system. Aggregation-prone, unstructured subunits cross the periplasmand are secreted through the outer membrane, after which they self-assemble.Here, significant progress is presented towards solving the high-resolutioncrystal structure of the novel amyloid transporter FapF fromPseudomonas,which facilitates the secretion of the amyloid-forming polypeptide FapC acrossthe bacterial outer membrane. This represents the first step towards obtainingstructural insight into the products of thePseudomonas  fapoperon. Initialattempts at crystallizing full-length and N-terminally truncated constructs byrefolding techniques were not successful; however, after preparing FapF106–430from the membrane fraction, reproducible crystals were obtained using thesitting-drop method of vapour diffusion. Diffraction data have been processedto 2.5 A resolution. These crystals belonged to the monoclinic space groupC121,with unit-cell parametersa= 143.4,b= 124.6,c= 80.4 A , = = 90, = 96.32 and three monomers in the asymmetric unit. It was found that the switch tocomplete detergent exchange into C8E4 was crucial for forming well diffractingcrystals, and it is suggested that this combined with limited proteolysis is apotentially useful protocol for membrane -barrel protein crystallography. Thethree-dimensional structure of FapF will provide invaluable information on themechanistic differences of biogenesis between the curli and Fap functionalamyloid systems.
AU  - Matthews,SJ
AU  - rouse,S
AU  - hawthorne
AU  - Lambert
AU  - hare
AU  - morgan,M
DO  - 10.1107/S2053230X16017921
EP  - 896
PY  - 2016///
SN  - 2053-230X
SP  - 892
TI  - Purification, crystallization and characterization of the Pseudomonas outer membrane protein FapF, a functional amyloid transporter
T2  - Acta Crystallographica Section F: Structural Biology Communications
UR  - http://dx.doi.org/10.1107/S2053230X16017921
UR  - http://hdl.handle.net/10044/1/42701
VL  - F72
ER  -
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