Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.

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Goshima N, Kawamura Y, Fukumoto A, Miura A, Honma R, Satoh R, Wakamatsu A, Yamamoto J, Kimura K, Nishikawa T, Andoh T, Iida Y, Ishikawa K, Ito E, Kagawa N, Kaminaga C, Kanehori K, Kawakami B, Kenmochi K, Kimura R, Kobayashi M, Kuroita T, Kuwayama H, Maruyama Y, Matsuo K, Minami K, Mitsubori M, Mori M, Morishita R, Murase A, Nishikawa A, Nishikawa S, Okamoto T, Sakagami N, Sakamoto Y, Sasaki Y, Seki T, Sono S, Sugiyama A, Sumiya T, Takayama T, Takayama Y, Takeda H, Togashi T, Yahata K, Yamada H, Yanagisawa Y, Endo Y, Imamoto F, Kisu Y, Tanaka S, Isogai T, Imai J, Watanabe S, Nomura N

Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.

Nat Methods. 2008 Dec;5(12):1011-7.

PubMed ID
19054851 [ View in PubMed
]
Abstract

Appropriate resources and expression technology necessary for human proteomics on a whole-proteome scale are being developed. We prepared a foundation for simple and efficient production of human proteins using the versatile Gateway vector system. We generated 33,275 human Gateway entry clones for protein synthesis, developed mRNA expression protocols for them and improved the wheat germ cell-free protein synthesis system. We applied this protein expression system to the in vitro expression of 13,364 human proteins and assessed their biological activity in two functional categories. Of the 75 tested phosphatases, 58 (77%) showed biological activity. Several cytokines containing disulfide bonds were produced in an active form in a nonreducing wheat germ cell-free expression system. We also manufactured protein microarrays by direct printing of unpurified in vitro-synthesized proteins and demonstrated their utility. Our 'human protein factory' infrastructure includes the resources and expression technology for in vitro proteome research.

DrugBank Data that Cites this Article

Polypeptides
NameUniProt ID
Histamine H2 receptorP25021Details
Vascular endothelial growth factor AP15692Details
RAC-alpha serine/threonine-protein kinaseP31749Details
5-hydroxytryptamine receptor 7P34969Details
Tyrosine-protein kinase FynP06241Details
Dipeptidyl peptidase 4P27487Details
Aquaporin-1P29972Details
Interferon betaP01574Details
Interferon gammaP01579Details
Peptidyl-prolyl cis-trans isomerase AP62937Details
Fibroblast growth factor 2P09038Details
ChymaseP23946Details
GTPase HRasP01112Details
Protein S100-A1P23297Details
Protein kinase C alpha typeP17252Details
Casein kinase II subunit alphaP68400Details
Caspase-7P55210Details
CalreticulinP27797Details
Proteasome subunit beta type-1P20618Details
Prostaglandin E2 receptor EP3 subtypeP43115Details
Casein kinase I isoform gamma-2P78368Details
Serine/threonine-protein kinase Chk1O14757Details
Bcl2-associated agonist of cell deathQ92934Details
G-protein coupled estrogen receptor 1Q99527Details
Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit gamma isoformQ13362Details
Mitogen-activated protein kinase 9P45984Details
Mitogen-activated protein kinase 8P45983Details
MAP kinase-activated protein kinase 3Q16644Details
Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoformQ08209Details
Adenylate kinase 2, mitochondrialP54819Details
Signal recognition particle 14 kDa proteinP37108Details
FollistatinP19883Details
Eukaryotic translation initiation factor 2 subunit 3P41091Details
Peroxiredoxin-1Q06830Details
Corticotropin-releasing factor receptor 1P34998Details
Calcium and integrin-binding protein 1Q99828Details
Cyclin-dependent kinase inhibitor 1P38936Details
Heterogeneous nuclear ribonucleoprotein KP61978Details
NucleophosminP06748Details
Probable ATP-dependent RNA helicase DDX5P17844Details
Myosin light chain kinase 3Q32MK0Details
Mitogen-activated protein kinase 6Q16659Details