Genetic studies in Arabidopsis implicate an α/ÎČ-hydrolase, KARRIKIN-INSENSITIVE 2 (KAI2) as a receptor for karrikins, germination-promoting butenolide small molecules found in the smoke of burned plants. However, direct biochemical evidence for the interaction between KAI2 and karrikin and for the mechanism of downstream signaling by a KAI2-karrikin complex remain elusive. We report crystallographic analyses and ligand-binding experiments for KAI2 recognition of karrikins. The karrikin-1 (KAR 1 ) ligand sits in the opening to the active site abutting a helical domain insert but distal from the canonical catalytic triad (Ser95-His246-Asp217) of α/ÎČ-hydrolases, consistent with the lack of detectable hydrolytic activity by purified KAI2. The closest approach of KAR 1 to Ser95-His246-Asp217 is 3.8 Ă
from His246. Six aromatic side chains, including His246, encapsulate KAR 1 through geometrically defined aromatic-aromatic interactions. KAR 1 binding induces a conformational change in KAI2 at the active site entrance. A crevice of hydrophobic residues linking the polar edge of KAR 1 and the helical domain insert suggests that KAI2-KAR 1 creates a contiguous interface for binding signaling partners in a liganddependent manner.plant signaling | wildfires | seed dormancy W ildfires recycle plant environments by releasing nutrients stored in plant tissues and often stimulate the germination of quiescent seeds (1). Germination-stimulating compounds, generated in the smoke of burning plants, include a seed dormancybreaking compound 3-methyl-2H-furo[2,3-c]pyran-2-one, a butenolide named karrikinolide-1 or karrikin-1 (KAR 1 ) (2). Previous reports reveal KAR 1 structure-activity relationships (SARs) and features of the physiological mode of action (3-7). Karrikinsinsensitive (kai) mutants in the model plant Arabidopsis thaliana identify some of the genetic components underlying karrikins' perception and signaling (8,9). Seeds of kai2 mutants are insensitive to KAR 1 -induced germination, suggesting that the KAI2 gene product is essential for KAR 1 perception and/or signal transduction (8, 9). KAI2, also known as HYPOSENSITIVE TO LIGHT (HTL) (10), is a member of the α/ÎČ-hydrolase superfamily commonly containing a catalytic triad, Ser(Cys)-His-Asp (11). Notably, several other members of the α/ÎČ-hydrolase superfamily sense, bind, and/or catalytically modify phytohormones. For example, the α/ÎČ-hydrolase SALICYLIC ACID-BINDING PROTEIN 2 (SABP2) converts methyl salicylate to salicylic acid through its esterase activity. The product of the reaction, salicylic acid, retains relatively high binding affinity to SABP2, which inhibits SABP2 catalytic activity and fine tunes salicylic acid levels (12). Another α/ÎČ-hydrolase family member GIBBERELLIN-INSENSITIVE DWARF 1 (GID1) lacks the highly conserved catalytic triad, but serves as the receptor of gibberellins (13,14). These previous studies have led to speculation that the KAI2 protein might be the receptor of karrikins (8,9). This hypothesis is supported by the recent discovery that ano...