Purification and primary structure determination of the N-terminal blocked protein, L11, from Escherichia coli ribosomes.
Protein L11 was isolated from the 50-S subunit of Escherichia coli ribosomes, using two salt extractions and two chromatographic separations on CM-cellulose. The unusual behavior of the protein when run on sodium dodecyl sulfate electrophoresis showed multiple bands. The complete primary structure of protein L11 is presented in detail. Its sequence was derived from peptides obtained by digesting the protein with trypsin, chymotrypsin, thermolysin, Staphylococcus aureus protease and, after modification, with trypsin. Chemical cleavage was performed with cyanogen bromide. Sequencing of the various peptides was achieved by manual micro-dansyl-Edman degradations and automatic methods. The N-terminal residue of the protein is blocked and was not degradable in the liquid-phase sequenator by the Edman method. It was identified by a combination of enzymatic cleavage and mass spectrometry. Protein L11 contain three methylated amino acid residues, a N alpha-trimethylalanine, and two residues of N epsilon-trimethyllysine. Their behaviour and influence in the sequence elucidation are described. The protein contains 141 amino acid residues and has a molecular weight of 14874. Secondary structure predictions of the protein are given, and its sequence is compared with those of other E. coli ribosomal proteins.
D000595:Amino Acid Sequence; D002268:Carboxypeptidases; D002918:Chymotrypsin; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000032:analysis*; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D012269:Ribosomal Proteins / Q000302:isolation & purification*; D012270:Ribosomes / Q000032:analysis*; D012697:Serine Endopeptidases; D013820:Thermolysin; D014357:Trypsin
D010446:Peptide Fragments; D012269:Ribosomal Proteins; C027624:ribosomal protein L11; D002268:Carboxypeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase; D014357:Trypsin; D013820:Thermolysin; D003488:Cyanogen Bromide