Conformationally constrained analogues of diacylglycerol (DAG). 28. DAG-dioxolanones reveal a new additional interaction site in the C1b domain of PKC delta.
Diacylglycerol (DAG) lactones have provided a powerful platform for structural exploration of the interactions between ligands and the C1 domains of protein kinase C (PKC). In this study, we report that DAG-dioxolanones, novel derivatives of DAG-lactones, exploit an additional point of contact (glutamine 27) in their binding with the C1b domain of PKC delta. Mutation of this point of contact to glutamate selectively impairs binding of the DAG-dioxolanones compared to that of the corresponding DAG-lactones (1200- to 3000-fold versus 35- to 55-fold, respectively). The differential response of this mutated C1b domain to the DAG-dioxolanones relative to the DAG-lactones provides a unique tool to probe the role of the C1b domain in PKC delta function, where the response to the DAG-lactones affords a positive control for retained function. Using this approach, we show that the C1b domain of PKC delta plays the predominant role in the translocation of PKC delta to the membrane in the presence of DAG.
Choi
Yongseok
Y
Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute-Frederick, National Institutes of Health, Frederick, Maryland 21702, USA.
Pu
Yongmei
Y
Peach
Megan L
ML
Kang
Ji-Hye
JH
Lewin
Nancy E
NE
Sigano
Dina M
DM
Garfield
Susan H
SH
Blumberg
Peter M
PM
Marquez
Victor E
VE
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004075:Diglycerides / Q000737:chemistry* / Q000494:pharmacology; D004148:Dioxolanes / Q000737:chemistry* / Q000494:pharmacology; D005973:Glutamine / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D007783:Lactones / Q000737:chemistry*; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D051745:Protein Kinase C-delta / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism
D004075:Diglycerides; D004148:Dioxolanes; D007783:Lactones; D011993:Recombinant Fusion Proteins; D005973:Glutamine; D049452:Green Fluorescent Proteins; D051745:Protein Kinase C-delta