Solution nuclear magnetic resonance structure of membrane-integral diacylglycerol kinase.
Escherichia coli diacylglycerol kinase (DAGK) represents a family of integral membrane enzymes that is unrelated to all other phosphotransferases. We have determined the three-dimensional structure of the DAGK homotrimer with the use of solution nuclear magnetic resonance. The third transmembrane helix from each subunit is domain-swapped with the first and second transmembrane segments from an adjacent subunit. Each of DAGK's three active sites resembles a portico. The cornice of the portico appears to be the determinant of DAGK's lipid substrate specificity and overhangs the site of phosphoryl transfer near the water-membrane interface. Mutations to cysteine that caused severe misfolding were located in or near the active site, indicating a high degree of overlap between sites responsible for folding and for catalysis.
Van Horn
Wade D
WD
Department of Biochemistry and Center for Structural Biology, Vanderbilt University, Nashville, TN 37232, USA.
Kim
Hak-Jun
HJ
Ellis
Charles D
CD
Hadziselimovic
Arina
A
Sulistijo
Endah S
ES
Karra
Murthy D
MD
Tian
Changlin
C
Sönnichsen
Frank D
FD
Sanders
Charles R
CR
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D055162:Biocatalysis; D020134:Catalytic Domain; D002462:Cell Membrane / Q000201:enzymology; D019852:Diacylglycerol Kinase / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017510:Protein Folding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D029968:Escherichia coli Proteins; D000255:Adenosine Triphosphate; D019852:Diacylglycerol Kinase