Nature Methods
- 4, 1019 - 1021 (2007)
Published online: 4 November 2007; | doi:10.1038/nmeth1118
In situ proteolysis for protein crystallization and structure determinationAiping Dong1, Xiaohui Xu2, Aled M Edwards1, 2 & Midwest Center for Structural Genomics(Changsoo Chang2, 3, Maksymilian Chruszcz2, 3, Marianne Cuff2, 3, Marcin Cymborowski2, 3, Rosa Di Leo2, 3, Olga Egorova2, 3, Elena Evdokimova2, 3, Ekaterina Filippova2, 3, Jun Gu2, 3, Jennifer Guthrie2, 3, Alexandr Ignatchenko2, 3, Andrzej Joachimiak2, 3, Natalie Klostermann2, 3, Youngchang Kim2, 3, Yuri Korniyenko2, 3, Wladek Minor2, 3, Qiuni Que2, 3, Alexei Savchenko2, 3, Tatiana Skarina2, 3, Kemin Tan2, 3, Alexander Yakunin2, 3, Adelinda Yee2, 3, Veronica Yim2, 3, Rongguang Zhang2, 3 & Hong Zheng2, 3) & Structural Genomics Consortium(Masato Akutsu1, 3, Cheryl Arrowsmith1, 3, George V Avvakumov1, 3, Alexey Bochkarev1, 3, Lars-Göran Dahlgren1, 3, Sirano Dhe-Paganon1, 3, Slav Dimov1, 3, Ludmila Dombrovski1, 3, Patrick Finerty Jr1, 3, Susanne Flodin1, 3, Alex Flores1, 3, Susanne Gräslund1, 3, Martin Hammerström1, 3, Maria Dolores Herman1, 3, Bum-Soo Hong1, 3, Raymond Hui1, 3, Ida Johansson1, 3, Yongson Liu1, 3, Martina Nilsson1, 3, Lyudmila Nedyalkova1, 3, Pär Nordlund1, 3, Tomas Nyman1, 3, Jinrong Min1, 3, Hui Ouyang1, 3, Hee-won Park1, 3, Chao Qi1, 3, Wael Rabeh1, 3, Limin Shen1, 3, Yang Shen1, 3, Deepthi Sukumard1, 3, Wolfram Tempel1, 3, Yufeng Tong1, 3, Lionel Tresagues1, 3, Masoud Vedadi1, 3, John R Walker1, 3, Johan Weigelt1, 3, Martin Welin1, 3, Hong Wu1, 3, Ting Xiao1, 3, Hong Zeng1, 3 & Haizhong Zhu1, 3)1
Structural Genomics Consortium, University of Toronto, 100 College Street, Toronto, Ontario M5G 1L5, Canada. 2
Midwest Center for Structural Genomics, University of Toronto, 112 College Street, Toronto, Ontario M5G 1L6, Canada. 3
Complete lists of authors appear at the end of this paper.
Correspondence should be addressed to Aled M Edwards aled.edwards@utoronto.ca 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.
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