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PMID: 17982461 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

In situ proteolysis for protein crystallization and structure determination.

Nature methods ·Vol. 4 ·No. 12 ·2007-12-00 ·Pages 1019-21

Dong A, Xu X, Edwards AM, Midwest Center for Structural Genomics, Structural Genomics Consortium, 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

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.

MeSH Terms
Crystallization/methods Crystallography/methods Peptide Hydrolases/chemistry Protein Conformation Proteins/chemistry,ultrastructure
Chemicals
Proteins Peptide Hydrolases
Authors & Affiliations
71 authors, click to expand affiliations / ORCID
Dong Aiping
Structural Genomics Consortium, University of Toronto, 100 College Street, Toronto, Ontario M5G 1L5, Canada.
Xu Xiaohui
Edwards Aled M
Midwest Center for Structural Genomics
Structural Genomics Consortium
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
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth1118
Published
2007-12-00
Epub
2007-00-04
Pages
1019-21
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC3366506
Subset
IM
Grants
NIGMS NIH HHS · U54 GM074942 · United States
NIGMS NIH HHS · U54 GM074942-02 · United States
Wellcome Trust · United Kingdom
NIGMS NIH HHS · GM074942 · United States
External Links
PubMed source DOI: 10.1038/nmeth1118 PMC Full Text View journal details
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