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Inactivation mechanism of the membrane protein diacylglycerol kinase in detergent solution
Author(s) -
Zhou Yufeng,
Lau Francis W.,
Nauli Sehat,
Yang Dawn,
Bowie James U.
Publication year - 2001
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.34201
Subject(s) - diacylglycerol kinase , chemistry , protein kinase c , biophysics , membrane , dissociation (chemistry) , protein kinase a , biochemistry , solubility , enzyme , diglyceride , protein aggregation , prkcq , biology , mitogen activated protein kinase kinase , organic chemistry
We have examined the irreversible inactivation mechanism of the membrane protein diacylglycerol kinase in the detergents n‐octyl‐β‐D‐glucopyranoside (OG) at 55°C and n‐decyl‐maltopyranoside (DM) at 80°C. Under no inactivation conditions did we find any direct evidence for the chemical modifications that are commonly found in soluble proteins. Moreover, protein inactivated at 55°C in OG could be reactivated by an unfolding and refolding protocol, suggesting that the protein is inactivated by a stable conformational change, not a covalent modification. We also found that the inactivation rate decreased with both increasing protein concentration and increasing thermodynamic stability, consistent with an inactivation pathway involving transient dissociation and/or unfolding of the protein. Our results suggest that the primary cause of diacylglycerol kinase inactivation is not low solubility, but poor intrinsic stability in the detergent environment.