Site-specific cleavage of the 40S ribosomal subunit reveals eukaryote-specific ribosomal protein S28 in the subunit head.
After resolving the crystal structure of the prokaryotic ribosome, mapping the proteins in the eukaryotic ribosome is a challenging task. We applied RNase H digestion to split the human 40S ribosomal subunit into head and body parts. Mass spectrometry of the proteins in the 40S subunit head revealed the presence of eukaryote-specific ribosomal protein S28e. Recombinant S28e was capable of specific binding to the 3' major domain of the 18S rRNA (K(a)=8.0±0.5×10(9) M(-1)). We conclude that S28e has a binding site on the 18S rRNA within the 40S subunit head.
Malygin
Alexey A
AA
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Karpova
Galina G
GG
D001665:Binding Sites; D056890:Eukaryota / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism*; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism*; D013045:Species Specificity
D012337:RNA, Ribosomal, 18S; C486166:RPS28 protein, human; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; D016914:Ribonuclease H