Identification of trimannoside-recognizing peptide sequences from a T7 phage display screen using a QCM device.
Here, we report on the identification of trimannoside-recognizing peptide sequences from a T7 phage display screen using a quartz-crystal microbalance (QCM) device. A trimannoside derivative that can form a self-assembled monolayer (SAM) was synthesized and used for immobilization on the gold electrode surface of a QCM sensor chip. After six sets of one-cycle affinity selection, T7 phage particles displaying PSVGLFTH (8-mer) and SVGLGLGFSTVNCF (14-mer) were found to be enriched at a rate of 17/44, 9/44, respectively, suggesting that these peptides specifically recognize trimannoside. Binding checks using the respective single T7 phage and synthetic peptide also confirmed the specific binding of these sequences to the trimannoside-SAM. Subsequent analysis revealed that these sequences correspond to part of the primary amino acid sequence found in many mannose- or hexose-related proteins. Taken together, these results demonstrate the effectiveness of our T7 phage display environment for affinity selection of binding peptides. We anticipate this screening result will also be extremely useful in the development of inhibitors or drug delivery systems targeting polysaccharides as well as further investigations into the function of carbohydrates in vivo.
Nishiyama
Kazusa
K
Department of Medicinal and Life Science, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Takakusagi
Yoichi
Y
Kusayanagi
Tomoe
T
Matsumoto
Yuki
Y
Habu
Shiori
S
Kuramochi
Kouji
K
Sugawara
Fumio
F
Sakaguchi
Kengo
K
Takahashi
Hideyo
H
Natsugari
Hideaki
H
Kobayashi
Susumu
S
D000595:Amino Acid Sequence; D017123:Bacteriophage T7; D001665:Binding Sites; D055808:Drug Discovery / Q000379:methods; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D008358:Mannose / Q000737:chemistry*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D014312:Trisaccharides / Q000737:chemistry*
D019151:Peptide Library; D010455:Peptides; D014312:Trisaccharides; D008358:Mannose