Structural genomics of the Thermotoga maritima proteome implemented in a high-throughput structure determination pipeline.
Structural genomics is emerging as a principal approach to define protein structure-function relationships. To apply this approach on a genomic scale, novel methods and technologies must be developed to determine large numbers of structures. We describe the design and implementation of a high-throughput structural genomics pipeline and its application to the proteome of the thermophilic bacterium Thermotoga maritima. By using this pipeline, we successfully cloned and attempted expression of 1,376 of the predicted 1,877 genes (73%) and have identified crystallization conditions for 432 proteins, comprising 23% of the T. maritima proteome. Representative structures from TM0423 glycerol dehydrogenase and TM0449 thymidylate synthase-complementing protein are presented as examples of final outputs from the pipeline.
Lesley
Scott A
SA
Joint Center for Structural Genomics, Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121, USA. lesley@gnf.org
Kuhn
Peter
P
Godzik
Adam
A
Deacon
Ashley M
AM
Mathews
Irimpan
I
Kreusch
Andreas
A
Spraggon
Glen
G
Klock
Heath E
HE
McMullan
Daniel
D
Shin
Tanya
T
Vincent
Juli
J
Robb
Alyssa
A
Brinen
Linda S
LS
Miller
Mitchell D
MD
McPhillips
Timothy M
TM
Miller
Mark A
MA
Scheibe
Daniel
D
Canaves
Jaume M
JM
Guda
Chittibabu
C
Jaroszewski
Lukasz
L
Selby
Thomas L
TL
Elsliger
Marc-Andre
MA
Wooley
John
J
Taylor
Susan S
SS
Hodgson
Keith O
KO
Wilson
Ian A
IA
Schultz
Peter G
PG
Stevens
Raymond C
RC
D003001:Cloning, Molecular; D016680:Genome, Bacterial; D008958:Models, Molecular; D016366:Open Reading Frames; D011487:Protein Conformation; D020543:Proteome; D020124:Thermotoga maritima / Q000235:genetics* / Q000378:metabolism
D020543:Proteome