Biochemical properties of the autophosphorylation of RLK5, a receptor like protein kinase from Arabidopsis thaliana英文文献.pdfVIP

BB Biochi~ic~a etBiophysicat~ta ELSEVIER Biochimica et Biophysica Acta 1208(1994) 65-74 Biochemical properties of the autophosphorylation of RLK5, a receptor-like protein kinase from Arabidopsis thaliana Mark A. Horn *, John C. Walker Division ofBiological Sciences, UniversityofMissouri-Columbia, Columbia,MO 65211, USA Received 11January 1994 Abstract The RLK5 gene of Arabidopsis thaliana encodes a novel receptor-like protein kinase. DNA sequence analysis suggests that the RLK5 protein contains an extracellular domain that has 21 tandemly repeated leucine-rich motifs linked, via a transmembrane hydrophobic region, to a protein kinase catalytic domain that is related to the serine/threonine family of protein kinases. To study the intrinsic biochemical properties of this protein kinase we have expressed the catalytic domain as two different recombinant fusion proteins in Escherichia coli. Both hybrid proteins have similar kinetic properties, autophosphorylate on serine and threonine residues and have s~gnificantly greater activity in the presence of Mn 2÷ than Mg 2÷. A lysine to glutamic acid substitution in the catalytic domain of RLK5 results in the catalytically inactive protein RLK5(Cat)I~711E. The active RLK5 protein can phosphorylate the inactive K711E protein and the K711E protein can par