Structure and dimerization of the kinase domain from yeast Snf1, a member of the Snf1/AMPK protein family.
The Snf1/AMPK kinases are intracellular energy sensors, and the AMPK pathway has been implicated in a variety of metabolic human disorders. Here we report the crystal structure of the kinase domain from yeast Snf1, revealing a bilobe kinase fold with greatest homology to cyclin-dependant kinase-2. Unexpectedly, the crystal structure also reveals a novel homodimer that we show also forms in solution, as demonstrated by equilibrium sedimentation, and in yeast cells, as shown by coimmunoprecipitation of differentially tagged intact Snf1. A mapping of sequence conservation suggests that dimer formation is a conserved feature of the Snf1/AMPK kinases. The conformation of the conserved alphaC helix, and the burial of the activation segment and substrate binding site within the dimer, suggests that it represents an inactive form of the kinase. Taken together, these studies suggest another layer of kinase regulation within the Snf1/AMPK family, and an avenue for development of AMPK-specific activating compounds.
Nayak
Vinod
V
The Wistar Institute, University of Pennsylvania, Philadelphia 19104, USA.
Zhao
Kehao
K
Wyce
Anastasia
A
Schwartz
Marc F
MF
Lo
Wan-Sheng
WS
Berger
Shelley L
SL
Marmorstein
Ronen
R
D000255:Adenosine Triphosphate / Q000737:chemistry; D000263:Adenylate Kinase / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D003461:Crystallography; D051357:Cyclin-Dependent Kinase 2 / Q000737:chemistry*; D019281:Dimerization; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry; D010770:Phosphotransferases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D012996:Solutions; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship; D015003:Yeasts / Q000737:chemistry / Q000235:genetics*
D009097:Multienzyme Complexes; D012996:Solutions; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D051357:Cyclin-Dependent Kinase 2; D000263:Adenylate Kinase