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 Que & 49 others Alexei 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

    Fingerprint

    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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