Immobilization of RNase S-Peptide on a single-stranded DNA-fixed gold surface and effective masking of its surface by an acridinyl poly(ethylene glycol).
Oligonucleotide-peptide conjugate was synthesized by coupling of RNase S-peptide to a 24-mer single-stranded DNA (ssDNA) oligonucleotide to be immobilized on its complementary ssDNA oligonucleotide-fixed gold surface of sensor chip or electrode. Immobilization of on the ssDNA-fixed gold surface through DNA duplex formation was confirmed by quartz crystal microbalance (QCM) and electrochemical measurements. After treating with a synthetic acridinyl poly(ethylene glycol) (APEG), specific interaction of S-protein with the S-peptide immobilized on the gold surface was demonstrated by QCM without nonspecific adsorption of unrelated proteins such as BSA and RNase A at the surfaces. This result suggested that the acridine parts of APEG could bind to the DNA duplex on the gold surface and the poly(ethylene glycol) parts were fastened on the surface to resist the adsorption of proteins. Thus, the combination of oligonucleotide-peptide conjugate, ssDNA-fixed chip and APEG with effective masking property provides a new tool for the analysis of specific peptide-protein interactions without disturbance by other unrelated proteins.
Ohtsuka
Keiichi
K
Department of Materials Science, Kyushu Institute of Technology, Kitakyushu-shi 804-8550, Japan.
Uemura
Keiko
K
Nojima
Takahiko
T
Waki
Michinori
M
Takenaka
Shigeori
S
D000166:Acridines / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation*; D004277:DNA, Single-Stranded; D006046:Gold; D020411:Oligonucleotide Array Sequence Analysis; D010446:Peptide Fragments / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D012260:Ribonucleases / Q000737:chemistry
D000166:Acridines; D004277:DNA, Single-Stranded; D010446:Peptide Fragments; C023560:ribonuclease S-peptide; D011092:Polyethylene Glycols; D006046:Gold; D012260:Ribonucleases