Identification of binding peptides of the ADAM15 disintegrin domain using phage display.
ADAM15 plays an important role in tumour development by interacting with integrins. In this study, we investigated the target peptides of the ADAM15 disintegrin domain. First, we successfully produced the recombinant human ADAM15 disintegrin domain (RADD) that could inhibit melanoma cell adhesion by using Escherichia coli. Second, four specific binding peptides (peptides A, B, C, and D) were selected using a phage display 12-mer peptide library. The screening protocol involved 4 rounds of positive panning on RADD and 2 rounds of subtractive selection with streptavidin. By using the BLAST software and a relevant protein database, integrin alpha v beta 3 was found to be homologous to peptide A. Synthetic peptide A had a highly inhibitory effect on RADD-integrin alpha v beta 3 binding. The results demonstrate the potential application of short peptides for disrupting high-affinity ADAM-integrin interactions.
Wu
Jing
J
School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, Jiangsu 214122, China.
Wu
Min-Chen
MC
Zhang
Lian-Fen
LF
Lei
Jian-Yong
JY
Feng
Lei
L
Jin
Jian
J
D051722:ADAM Proteins / Q000737:chemistry / Q000502:physiology*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D008954:Models, Biological; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D040901:Proteomics / Q000379:methods
D016023:Integrins; D008565:Membrane Proteins; D019151:Peptide Library; D010455:Peptides; D051722:ADAM Proteins; C098660:ADAM15 protein, human