Binding of brassinosteroids to the extracellular domain of plant receptor kinase BRI1.
Both animals and plants use steroids as signalling molecules during growth and development. Animal steroids are principally recognized by members of the nuclear receptor superfamily of transcription factors. In plants, BRI1, a leucine-rich repeat (LRR) receptor kinase localized to the plasma membrane, is a critical component of a receptor complex for brassinosteroids. Here, we present the first evidence for direct binding of active brassinosteroids to BRI1 using a biotin-tagged photoaffinity castasterone (BPCS), a biosynthetic precursor of brassinolide (the most active of the brassinosteroids). Binding studies using BPCS, (3)H-labelled brassinolide and recombinant BRI1 fragments show that the minimal binding domain for brassinosteroids consists of a 70-amino acid island domain (ID) located between LRR21 and LRR22 in the extracellular domain of BRI1, together with the carboxy-terminal flanking LRR (ID-LRR22). Our results demonstrate that brassinosteroids bind directly to the 94 amino acids comprising ID-LRR22 in the extracellular domain of BRI1, and define a new binding domain for steroid hormones.
Kinoshita
Toshinori
T
Howard Hughes Medical Institute and Plant Biology Laboratory, The Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, California 92037, USA.
Caño-Delgado
Ana
A
Seto
Hideharu
H
Hiranuma
Sayoko
S
Fujioka
Shozo
S
Yoshida
Shigeo
S
Chory
Joanne
J
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism; D060406:Brassinosteroids; D002777:Cholestanols / Q000378:metabolism*; D003432:Cross-Linking Reagents; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011485:Protein Binding; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013260:Steroids, Heterocyclic / Q000378:metabolism*
D029681:Arabidopsis Proteins; D060406:Brassinosteroids; D002777:Cholestanols; D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D013260:Steroids, Heterocyclic; D001710:Biotin; C083545:castasterone; D011494:Protein Kinases; C108270:BRI1 protein, Arabidopsis; C023623:brassinolide