In situ proteolysis for protein crystallization and structure determination

Aiping Dong, Xiaohui Xu, Aled M. Edwards, Changsoo Chang, Maksymilian Chruszcz, Marianne Cuff, Marcin Cymborowski, Rosa Di Leo, Olga Egorova, Elena Evdokimova, Ekaterina Filippova, Jun Gu, Jennifer Guthrie, Alexandr Ignatchenko, Andrzej Joachimiak, Natalie Klostermann, Youngchang Kim, Yuri Korniyenko, Wladek Minor, Qiuni QueAlexei Savchenko, Tatiana Skarina, Kemin Tan, Alexander Yakunin, Adelinda Yee, Veronica Yim, Rongguang Zhang, Hong Zheng, Masato Akutsu, Cheryl Arrowsmith, George V. Avvakumov, Alexey Bochkarev, Lars Göran Dahlgren, Sirano Dhe-Paganon, Slav Dimov, Ludmila Dombrovski, Patrick Finerty, Susanne Flodin, Alex Flores, Susanne Gräslund, Martin Hammerström, Maria Dolores Herman, Bum Soo Hong, Raymond Hui, Ida Johansson, Yongson Liu, Martina Nilsson, Lyudmila Nedyalkova, Pär Nordlund, Tomas Nyman, Jinrong Min, Hui Ouyang, Hee Won Park, Chao Qi, Wael Rabeh, Limin Shen, Yang Shen, Deepthi Sukumard, Wolfram Tempel, Yufeng Tong, Lionel Tresagues, Masoud Vedadi, John R. Walker, Johan Weigelt, Martin Welin, Hong Wu, Ting Xiao, Hong Zeng, Haizhong Zhu

Research output: Contribution to journalArticle

Abstract

We tested the general applicability of in situ proteolysis to form protein crystals suitable for structure determination by adding a protease (chymotrypsin or trypsin) digestion step to crystallization trials of 55 bacterial and 14 human proteins that had proven recalcitrant to our best efforts at crystallization or structure determination. This is a work in progress; so far we determined structures of 9 bacterial proteins and the human aminoimidazole ribonucleotide synthetase (AIRS) domain.

Original languageEnglish (US)
Pages (from-to)1019-1021
Number of pages3
JournalNature Methods
Volume4
Issue number12
DOIs
StatePublished - Dec 1 2007

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Proteolysis
Crystallization
Ribonucleotides
Bacterial Proteins
Chymotrypsin
Ligases
Trypsin
Digestion
Proteins
Peptide Hydrolases
Crystals
trypsin drug combination chymotrypsin

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Dong, A., Xu, X., Edwards, A. M., Chang, C., Chruszcz, M., Cuff, M., ... Zhu, H. (2007). In situ proteolysis for protein crystallization and structure determination. Nature Methods, 4(12), 1019-1021. https://doi.org/10.1038/nmeth1118

In situ proteolysis for protein crystallization and structure determination. / Dong, Aiping; Xu, Xiaohui; Edwards, Aled M.; Chang, Changsoo; Chruszcz, Maksymilian; Cuff, Marianne; Cymborowski, Marcin; Di Leo, Rosa; Egorova, Olga; Evdokimova, Elena; Filippova, Ekaterina; Gu, Jun; Guthrie, Jennifer; Ignatchenko, Alexandr; Joachimiak, Andrzej; Klostermann, Natalie; Kim, Youngchang; Korniyenko, Yuri; Minor, Wladek; Que, Qiuni; Savchenko, Alexei; Skarina, Tatiana; Tan, Kemin; Yakunin, Alexander; Yee, Adelinda; Yim, Veronica; Zhang, Rongguang; Zheng, Hong; Akutsu, Masato; Arrowsmith, Cheryl; Avvakumov, George V.; Bochkarev, Alexey; Dahlgren, Lars Göran; Dhe-Paganon, Sirano; Dimov, Slav; Dombrovski, Ludmila; Finerty, Patrick; Flodin, Susanne; Flores, Alex; Gräslund, Susanne; Hammerström, Martin; Herman, Maria Dolores; Hong, Bum Soo; Hui, Raymond; Johansson, Ida; Liu, Yongson; Nilsson, Martina; Nedyalkova, Lyudmila; Nordlund, Pär; Nyman, Tomas; Min, Jinrong; Ouyang, Hui; Park, Hee Won; Qi, Chao; Rabeh, Wael; Shen, Limin; Shen, Yang; Sukumard, Deepthi; Tempel, Wolfram; Tong, Yufeng; Tresagues, Lionel; Vedadi, Masoud; Walker, John R.; Weigelt, Johan; Welin, Martin; Wu, Hong; Xiao, Ting; Zeng, Hong; Zhu, Haizhong.

In: Nature Methods, Vol. 4, No. 12, 01.12.2007, p. 1019-1021.

Research output: Contribution to journalArticle

Dong, A, Xu, X, Edwards, AM, Chang, C, Chruszcz, M, Cuff, M, Cymborowski, M, Di Leo, R, Egorova, O, Evdokimova, E, Filippova, E, Gu, J, Guthrie, J, Ignatchenko, A, Joachimiak, A, Klostermann, N, Kim, Y, Korniyenko, Y, Minor, W, Que, Q, Savchenko, A, Skarina, T, Tan, K, Yakunin, A, Yee, A, Yim, V, Zhang, R, Zheng, H, Akutsu, M, Arrowsmith, C, Avvakumov, GV, Bochkarev, A, Dahlgren, LG, Dhe-Paganon, S, Dimov, S, Dombrovski, L, Finerty, P, Flodin, S, Flores, A, Gräslund, S, Hammerström, M, Herman, MD, Hong, BS, Hui, R, Johansson, I, Liu, Y, Nilsson, M, Nedyalkova, L, Nordlund, P, Nyman, T, Min, J, Ouyang, H, Park, HW, Qi, C, Rabeh, W, Shen, L, Shen, Y, Sukumard, D, Tempel, W, Tong, Y, Tresagues, L, Vedadi, M, Walker, JR, Weigelt, J, Welin, M, Wu, H, Xiao, T, Zeng, H & Zhu, H 2007, 'In situ proteolysis for protein crystallization and structure determination', Nature Methods, vol. 4, no. 12, pp. 1019-1021. https://doi.org/10.1038/nmeth1118
Dong A, Xu X, Edwards AM, Chang C, Chruszcz M, Cuff M et al. In situ proteolysis for protein crystallization and structure determination. Nature Methods. 2007 Dec 1;4(12):1019-1021. https://doi.org/10.1038/nmeth1118
Dong, Aiping ; Xu, Xiaohui ; Edwards, Aled M. ; Chang, Changsoo ; Chruszcz, Maksymilian ; Cuff, Marianne ; Cymborowski, Marcin ; Di Leo, Rosa ; Egorova, Olga ; Evdokimova, Elena ; Filippova, Ekaterina ; Gu, Jun ; Guthrie, Jennifer ; Ignatchenko, Alexandr ; Joachimiak, Andrzej ; Klostermann, Natalie ; Kim, Youngchang ; Korniyenko, Yuri ; Minor, Wladek ; Que, Qiuni ; Savchenko, Alexei ; Skarina, Tatiana ; Tan, Kemin ; Yakunin, Alexander ; Yee, Adelinda ; Yim, Veronica ; Zhang, Rongguang ; Zheng, Hong ; Akutsu, Masato ; Arrowsmith, Cheryl ; Avvakumov, George V. ; Bochkarev, Alexey ; Dahlgren, Lars Göran ; Dhe-Paganon, Sirano ; Dimov, Slav ; Dombrovski, Ludmila ; Finerty, Patrick ; Flodin, Susanne ; Flores, Alex ; Gräslund, Susanne ; Hammerström, Martin ; Herman, Maria Dolores ; Hong, Bum Soo ; Hui, Raymond ; Johansson, Ida ; Liu, Yongson ; Nilsson, Martina ; Nedyalkova, Lyudmila ; Nordlund, Pär ; Nyman, Tomas ; Min, Jinrong ; Ouyang, Hui ; Park, Hee Won ; Qi, Chao ; Rabeh, Wael ; Shen, Limin ; Shen, Yang ; Sukumard, Deepthi ; Tempel, Wolfram ; Tong, Yufeng ; Tresagues, Lionel ; Vedadi, Masoud ; Walker, John R. ; Weigelt, Johan ; Welin, Martin ; Wu, Hong ; Xiao, Ting ; Zeng, Hong ; Zhu, Haizhong. / In situ proteolysis for protein crystallization and structure determination. In: Nature Methods. 2007 ; Vol. 4, No. 12. pp. 1019-1021.
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