Open Access`Seeding' with protease to optimize protein crystallization conditions in in situ proteolysis
Author(s) -
Huang Jinguang,
Gong Yanmei,
Huang Dan,
Haire Lesley,
Liu Junfeng,
Peng Youliang
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911201161x
Subject(s) - proteolysis , crystallization , protease , seeding , in situ , protein crystallization , chemistry , materials science , biochemistry , enzyme , biology , organic chemistry , agronomy
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.