Expression, purification and crystallization of the ammonium transporter Amt-1 from Archaeoglobus fulgidus

被引:10
作者
Andrade, SLA [1 ]
Dickmanns, A [1 ]
Ficner, R [1 ]
Einsle, O [1 ]
机构
[1] Univ Gottingen, Inst Mikrobiol & Genet, Abt Mol Strukt Biol, D-37077 Gottingen, Germany
来源
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2005年 / 61卷
关键词
D O I
10.1107/S1744309105027004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Ammonium transporters (Amts) are a class of membrane-integral transport proteins found in organisms from all kingdoms of life. Their key function is the transport of nitrogen in its reduced bioavailable form, ammonia, across cellular membranes, a crucial step in nitrogen assimilation for biosynthetic purposes. The genome of the hyperthermophilic archaeon Archaeoglobus fulgidus has been annotated with three individual genes for ammonium transporters, amt1-3, the roles of which are as yet unknown. The amt1 gene product has been produced by heterologous overexpression in Escherichia coli and the resulting protein has been purified to electrophoretic homogeneity. Crystals of Amt-1 have been obtained by sitting-drop vapour diffusion and diffraction data have been collected.
引用
收藏
页码:861 / 863
页数:3
相关论文
共 17 条
[1]   Purification of the Escherichia coli ammonium transporter AmtB reveals a trimeric stoichiometry [J].
Blakey, D ;
Leech, A ;
Thomas, GH ;
Coutts, G ;
Findlay, K ;
Merrick, M .
BIOCHEMICAL JOURNAL, 2002, 364 :527-535
[2]   Antagonism of PII signalling by the AmtB protein of Escherichia coli [J].
Blauwkamp, TA ;
Ninfa, AJ .
MOLECULAR MICROBIOLOGY, 2003, 48 (04) :1017-1028
[3]   Electron and atomic force microscopy of the trimeric ammonium transporter AmtB [J].
Conroy, MJ ;
Jamieson, SJ ;
Blakey, D ;
Kaufmann, T ;
Engel, A ;
Fotiadis, D ;
Merrick, M ;
Bullough, PA .
EMBO REPORTS, 2004, 5 (12) :1153-1158
[4]   Membrane sequestration of the signal transduction protein GlnK by the ammonium transporter AmtB [J].
Coutts, G ;
Thomas, G ;
Blakey, D ;
Merrick, M .
EMBO JOURNAL, 2002, 21 (04) :536-545
[5]   The Rh complex exports ammonium from human red blood cells [J].
Hemker, MB ;
Cheroutre, G ;
van Zwieten, R ;
Maaskant-van Wijk, PA ;
Roos, D ;
Loos, JA ;
van der Schoot, CE ;
von dem Borne, AEGK .
BRITISH JOURNAL OF HAEMATOLOGY, 2003, 122 (02) :333-340
[6]   Ammonium sensing in Escherichia coli -: Role of the ammonium transporter AmtB and AmtB-GlnK complex formation [J].
Javelle, A ;
Severi, E ;
Thornton, J ;
Merrick, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (10) :8530-8538
[7]   Mechanism of ammonia transport by Amt/MEP/Rh:: Structure of AmtB at 1.3.5 Å [J].
Khademi, S ;
O'Connell, J ;
Remis, J ;
Robles-Colmenares, Y ;
Miericke, LJW ;
Stroud, RM .
SCIENCE, 2004, 305 (5690) :1587-1594
[8]   BACTERIAL AMMONIUM TRANSPORT [J].
KLEINER, D .
FEMS MICROBIOLOGY LETTERS, 1985, 32 (02) :87-100
[9]   The MEP2 ammonium permease regulates pseudohyphal differentiation in Saccharomyces cerevisiae [J].
Lorenz, MC ;
Heitman, J .
EMBO JOURNAL, 1998, 17 (05) :1236-1247
[10]   Uniport of NH4+ by the root hair plasma membrane ammonium transporter LeAMT1;1 [J].
Ludewig, U ;
von Wirén, N ;
Frommer, WB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (16) :13548-13555