Specific Ser-Pro phosphorylation by the RNA-recognition motif containing kinase KIS.
We present here a first appraisal of the phosphorylation site specificity of KIS (for 'kinase interacting with stathmin'), a novel mammalian kinase that has the unique feature among kinases to possess an RNP type RNA-recognition motif (RRM). In vitro kinase assays using various standard substrates revealed that KIS has a narrow specificity, with myelin basic protein (MBP) and synapsin I being the best in vitro substrates among those tested. Mass spectrometry and peptide sequencing allowed us to identify serine 164 of MBP as the unique site phosphorylated by KIS. Phosphorylation of synthetic peptides indicated the importance of the proline residue at position +1. We also identified a tryptic peptide of synapsin I phosphorylated by KIS and containing a phosphorylatable Ser-Pro motif. Altogether, our results suggest that KIS preferentially phosphorylates proline directed residues but has a specificity different from that of MAP kinases and cdks.
Maucuer
A
A
INSERM U440, Institut du Fer à Moulin, Paris, France. maucer@ifm.inserm.fr
Le Caer
J P
JP
Manceau
V
V
Sobel
A
A
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D047908:Intracellular Signaling Peptides and Proteins; D013058:Mass Spectrometry; D004676:Myelin Basic Protein / Q000378:metabolism; D010766:Phosphorylation; D011392:Proline / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D012694:Serine / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D016704:Synapsins / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D013997:Time Factors
D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D004676:Myelin Basic Protein; D011994:Recombinant Proteins; D016704:Synapsins; D013912:Threonine; D012694:Serine; D012313:RNA; D011392:Proline; D017346:Protein Serine-Threonine Kinases