Open AccessThe Primary Structure of the Coat Protein of the Broad‐Host‐Range RNA Bacteriophage PRR1
Author(s) -
DHAESE Patrick,
VANDEKERCKHOVE Joël S.,
MONTAGU Marc C.
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb12903.x
Subject(s) - biology , cyanogen bromide , rna , biochemistry , edman degradation , peptide sequence , protease , amino acid , bacteriophage , trypsin , thermolysin , escherichia coli , enzyme , gene
The complete amino acid sequence of the coat protein of RNA bacteriophage PRR1 is presented. After thermolysin digestion, 26 peptides were isolated, covering the complete coat protein chain. Their alignment was established in part using automated Edman degradation on the intact protein, in part with overlapping peptides obtained by enzymic hydrolysis with trypsin, pepsin, subtilisin and Staphylococcus aureus protease, and by chemical cleavage with cyanogen bromide and N ‐bromo‐succinimide. To obtain the final overlaps, a highly hydrophobic, insoluble tryptic peptide was sequenced for seven steps by the currently used manual dansyl‐Edman degradation procedure, which was slightly modified for application on insoluble peptides. PRR1 coat protein contains 131 amino acids, corresponding to molecular weight of 14534. It is highly hydrophobic, and the residues with ionizable side chains are distributed unevenly: acidic residues are absent in the middle third of the sequence, whereas a clustering of basic residues occurs between positions 44 and 62. PRR1 coat protein was compared with the coat proteins of RNA coliphages MS2 and Qβ, and the minimum mutation distance was calculated for both comparisons. It is highly probable that PRR1, Qβ and MS2 share a common ancestor. The basic region present in the three coat proteins is recognized as an essential structural feature of RNA phage coat proteins.