'Seeding' with protease to optimize protein crystallization conditions in in situ proteolysis.
In situ proteolysis is one of the most effective rescue strategies for protein crystallization, and optimization of the ratio between the protein and the protease is one of the key steps in the process. Seeding is a very powerful tool to optimize crystallization conditions and can be performed by most crystallization robots. Addition of protease instead of seed stock using a robot can be used to optimize the concentration of protease in in situ proteolysis experiments and has been successfully tested using two proteins.
State Key Laboratory of Agribiotechnology and MOA Key Laboratory of Plant Pathology, China Agricultural University, 2 Yuanmingyuan Xilu, Beijing 100193, People's Republic of China.
Jinguang
J
Huang
Yanmei
Y
Gong
Dan
D
Huang
Lesley
L
Haire
Junfeng
J
Liu
Youliang
Y
Peng
D018834:Chaperonin 60 / Q000032:analysis* / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry
D018834:Chaperonin 60; D010450:Endopeptidases