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.
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
Scott A
SA
Lesley
Peter
P
Kuhn
Adam
A
Godzik
Ashley M
AM
Deacon
Irimpan
I
Mathews
Andreas
A
Kreusch
Glen
G
Spraggon
Heath E
HE
Klock
Daniel
D
McMullan
Tanya
T
Shin
Juli
J
Vincent
Alyssa
A
Robb
Linda S
LS
Brinen
Mitchell D
MD
Miller
Timothy M
TM
McPhillips
Mark A
MA
Miller
Daniel
D
Scheibe
Jaume M
JM
Canaves
Chittibabu
C
Guda
Lukasz
L
Jaroszewski
Thomas L
TL
Selby
Marc-Andre
MA
Elsliger
John
J
Wooley
Susan S
SS
Taylor
Keith O
KO
Hodgson
Ian A
IA
Wilson
Peter G
PG
Schultz
Raymond C
RC
Stevens
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