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In situ proteolysis for protein crystallization and structure determination.

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.

SUBMITTER: Dong A 

PROVIDER: S-EPMC3366506 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature

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In situ proteolysis for protein crystallization and structure determination.

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

Nature methods 20071104 12

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. ...[more]

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