Cloning, expression, purification, crystallization and preliminary X-ray diffraction of a lysine-specific permease from Pseudomonas aeruginosa
Citation:
Nji, E. Li, D. Doyle, D.A. Caffrey, M., Cloning, expression, purification, crystallization and preliminary X-ray diffraction of a lysine-specific permease from Pseudomonas aeruginosa, Acta Crystallographica Section F Structural Biology Communications, 70, 2014, 1362 - 1367Download Item:
Abstract:
The prokaryotic lysine-specific permease (LysP) belongs to the amino acid–
polyamine–organocation (APC) transporter superfamily. In the cell, members
of this family are responsible for the uptake and recycling of nutrients, for
the maintenance of a constant internal ion concentration and for cell volume
regulation. The detailed mechanism of substrate selectivity and transport of
l
-lysine by LysP is not understood. A high-resolution crystal structure would
enormously facilitate such an understanding. To this end, LysP from
Pseudomonas aeruginosa
was recombinantly expressed in
Escherichia coli
and
purified to near homogeneity by immobilized metal ion-affinity chromatography
(IMAC) and size-exclusion chromatography (SEC). Hexagonal- and rod-shaped
crystals were obtained in the presence of
l
-lysine and the
l
-lysine analogue
l
-4-
thialysine by vapour diffusion and diffracted to 7.5 A
̊
resolution. The diffraction
data were indexed in space group
P
2
1
, with unit-cell parameters
a
= 169.53,
b
= 169.53,
c
= 290.13 A
̊
,
= 120
Sponsor
Grant Number
Science Foundation Ireland (SFI)
12/IA/125
Author's Homepage:
http://people.tcd.ie/mcaffreDescription:
PUBLISHED
Author: CAFFREY, MARTIN
Type of material:
Journal ArticleCollections
Series/Report no:
Acta Crystallographica Section F Structural Biology Communications70
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Full text availableKeywords:
amino acid– polyamine–organocation (APC)DOI:
http://dx.doi.org/10.1107/S2053230X14017865Metadata
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