Dipeptide seryl-histidine and related oligopeptides cleave DNA, protein, and a carboxyl ester.
The amino acids histidine (His) and serine (Ser), or amino acids similar to Ser, function together as key catalytic amino acids in the active sites of such diverse enzymes as the serine- and thiol-proteases, lipases, and esterases. Ser and His are also conserved in the intein-extein junctions of the phylogenetically widespread self-splicing proteins and at the N- and C-termini of the homing endonucleases spliced from them. Here we show that the dipeptide seryl-histidine (Ser-His) and related oligopeptides can themselves cleave DNA, protein, and the ester p-nitrophenyl acetate (p-NPA) over wide ranges of pH and temperature. Denaturing polyacrylamide gel electrophoresis (PAGE) of 5'-end labeled DNA samples incubated with Ser-His reveals a pattern of two bands per nucleotide position, consistent with the generation of both 3'-hydroxyl and 3'-phosphate DNA cleavage fragments, as would be expected of phosphodiester hydrolysis by Ser-His. To the best of our knowledge, Ser-His is the shortest peptide ever reported to show cleavage activity with multiple categories of natural substrates. The amenability of the dipeptide to variation through addition of amino acid residues, either internally or to the C-terminus while retaining its multiple cleavage activities, combined with its reactivity over wide ranges of pH and temperature, demonstrates the evolutionary capacity of the Ser/His dyad and evokes many questions about possible roles it may have played in molecular evolution and its potential role as a core for selection of oligopeptides with enhanced cleavage activities and target specificity.
Li
Y
Y
Edison Biotechnology Institute of Ohio University, Athens 45701, USA.
Zhao
Y
Y
Hatfield
S
S
Wan
R
R
Zhu
Q
Q
Li
X
X
McMills
M
M
Ma
Y
Y
Li
J
J
Brown
K L
KL
He
C
C
Liu
F
F
Chen
X
X
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002417:Cattle; D004279:DNA, Viral / Q000378:metabolism*; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism*; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism; D019143:Evolution, Molecular; D066298:In Vitro Techniques; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013379:Substrate Specificity
D004279:DNA, Viral; D004151:Dipeptides; D004798:Enzymes; D009842:Oligopeptides; C422849:seryl-histidine; D012710:Serum Albumin, Bovine