Exploring some of the physico-chemical properties of the LAMMER protein kinase DOA of Drosophila.
Members of the highly conserved LAMMER family of protein kinases have been described in all eukaryotes. LAMMER kinases possess markedly similar peptide motifs in their kinase catalytic subdomains that are responsible for phosphotransfer and substrate interaction, suggesting that family members serve similar functions in widely diverged species. This hypothesis is supported by their phosphorylation of SR and SR-related proteins in diverged species. Here we describe a 3-dimensional homology model of the catalytic domain of DOA, a representative LAMMER kinase, encoded by the Drosophila locus Darkener of apricot (Doa). Homology modeling of DOA based on a Sky1p template revealed a highly conserved structural framework within conserved core regions. These adopt typical kinase folding like that of other protein kinases. However, in contrast to Sky1p, some structural features, such as those in helix alphaC suggest that the DOA kinase is not a constitutively active enzyme but requires activation. This may occur by phosphorylation within an activation loop that forms a broad turn and in which interactions between the side chains occur across the loop. The fold of the activation loop is stabilized through interactions with residues in the C-terminal tail, which is not part of the conserved kinase core and is variable among protein kinases. Immediately following the activation loop in the segment between the beta9 sheet and helix alphaF is a P + 1 loop. The electrostatic surface potential of the DOA substrate-binding groove is largely negative, as it is in other known SR protein kinases, suggesting that DOA substrates must be basic. All differences between D. melanogaster and other Drosophila species are single amino acid changes situated in regions outside of any alpha-helices or beta-sheets, and after modeling these had absolutely no visible effect on protein structure. The absence of evolved amino acid changes among 12 Drosophila species that would cause at least predictable changes in DOA structure indicate that evolution has already selected evolved mutations for having minimal effect on kinase structure.
Farkas
Robert
R
Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia. ueenfark@savba.savba.sk
Kováciková
Michaela
M
Liszeková
Denisa
D
Beno
Milan
M
Danis
Peter
P
Rabinow
Leonard
L
Chase
Bruce A
BA
Raska
Ivan
I
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D029721:Drosophila Proteins / Q000737:chemistry* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000201:enzymology*; D008958:Models, Molecular; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D016415:Sequence Alignment
D029721:Drosophila Proteins; D029701:Saccharomyces cerevisiae Proteins; C089977:Doa protein, Drosophila; C490216:SKY1 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases