The use of hydroxylamine cleavage to produce a fragment of ribosomal protein S4 which retains the capacity to specifically bind 16S ribosomal RNA.
In previous reports we have described the isolation of fragments of 30S ribosomal protein S4 using a number of different enzymatic and chemical cleavage techniques. These experiments were designed to determine the region of the protein responsible for 16S RNA recognition. We report here the isolation of two fragments produced by the hydroxylamine cleavage of the asparaginyl-glycyl peptide bond between positions 124 and 125. The purified fragments were chemically identified and tested for RNA binding capacity. The fragment consisting of residues 1-124 retains RNA binding activity and the fragment 125-203 is totally without RNA binding function. These results and previous results strongly suggest that the domain of protein S4 responsible for 16S RNA specific association is within the region consisting of residues 46-124.
D001665:Binding Sites; D004926:Escherichia coli; D019811:Hydroxylamine; D006898:Hydroxylamines; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D006898:Hydroxylamines; D010446:Peptide Fragments; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D019811:Hydroxylamine