A Protein Factory For High Throughput Structural Biology

Protein Production UK (PPUK) is a new initiative in the Research Complex that addresses sample preparation for challenging proteins and multicomponent complexes. PPUK will be part of the Rosalind Franklin Institute and closely linked to the Central facilities on the Harwell campus. PPUK aims to develop new and innovative methods and to work in partnership with academic groups and industry to enable uptake of the technology.

PPUK has an established pipeline for protein production based on the technology developed by the Oxford Protein Production Facility, involving construction of multi-purpose vectors (e.g. for producing complexes, and membrane proteins), high-throughput expression screening in different cell hosts (microbial, insect and mammalian cells) combined with scale-up to multi-litre culture volumes and downstream processing. New technology platforms that are under development include production of single-domain antibodies for the stabilisation of membrane proteins for analysis by cryo-EM.

©2010 Medical Research Council

To see the protein production line in action and how we crystallize proteins, please watch the video below

 

 

 

Recent papers

Ahmed AR, Owens RJ,  Stubbs CD, Parker AW, Hitchman R, Yadav RB, Dumoux M, Hawes C,  & Botchway SW. Direct imaging of the recruitmentand phosphorylation of S6K1 in the mTORC1 pathway in living cells Scientific Reports  2019) 9:3408 doi.org/10.1038/s41598-019-39410-z.

Romanello L, Zeraik AE, de Freitas Fernandes A, Torini JR, Bird LE, Nettleship JE, Rada H, Reddivari Y, Owens RJ, Serrão VHB, DeMarco R, Brandão-Neto J, Pereira HD. In vitro and in vivo characterization of the multiple isoforms of Schistosoma mansoni Hypoxanthine-guanine phosphoribosyltransferases. Mol Biochem Parasitol. 2019 Feb 14. pii: S0166-6851(18)30243-3. doi: 10.1016/j.molbiopara.2019.02.005.

Sabaratnam K, Renner M, Paesen G, Harlos K, Nair V, Owens RJ, Grimes JM. Insights from the crystal structure of the chicken CREB3 bZIP suggest members of the CREB3 subfamily transcription factors may be activated in response to oxidative stress. Protein Sci. 2019 Jan 17. doi: 10.1002/pro.3573.

Kim YC, Lopez-Camacho C, Nettleship JE, Rahman N, Hill ML, Silva-Reyes L, Ortiz-Martinez G, Figueroa-Aguilar G, Mar MA, Vivanco-Cid H, Rollier CS, Zitzmann N, Viveros-Sandoval ME, Owens RJ, Reyes-Sandoval A. Optimization of Zika virus envelope protein production for ELISA and correlation of antibody titers with virus neutralization in Mexican patients from an arbovirus endemic region. Virol J. 2018 Dec 27;15(1):193.

Salimraj R, Hinchliffe P, Kosmopoulou M, Tyrrell JM, Brem J, van Berkel SS, Verma A, Owens RJ, McDonough MA, Walsh TR, Schofield CJ, Spencer J. Crystal Structures of VIM-1 Complexes Explain Active Site Heterogeneity in VIM-Class Metallo-β-Lactamases. FEBS J. 2018 Nov 15. doi: 10.1111/febs.14695.

Torini JR, Romanello L, Batista FAH, Serrão VHB, Faheem M, Zeraik AE, Bird L, Nettleship J, Reddivari Y, Owens R, DeMarco R, Borges JC, Brandão-Neto J, Pereira HD. The molecular structure of Schistosoma mansoni PNP isoform 2 provides insights into the nucleoside selectivity of PNPs. PLoS One. 2018 Sep 7;13(9):e0203532. doi: 10.1371/journal.pone.0203532.

Yogavel M, Nettleship JE, Sharma A, Harlos K, Jamwal A, Chaturvedi R, Sharma M, Jain V, Chhibber-Goel J, Sharma A. Structure of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase-dihydropteroate synthase from Plasmodium vivax sheds light on drug resistance. J Biol Chem. 2018 Aug 13. pii: jbc.RA118.00455

Trundová M, Kovaľ T, Owens RJ, Fejfarová K, Dušková J, Kolenko P, Dohnálek J. Highly stable single-strand-specific 3'-nuclease/nucleotidase from Legionella pneumophila. Int J Biol Macromol. 2018 Jul 15;114:776-787.

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