Skip to main content

Advertisement

SpringerLink
Log in

Identification of binding peptides of the ADAM15 disintegrin domain using phage display

  • Published:
Journal of Biosciences Aims and scope Submit manuscript

Abstract

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 α ν β 3 was found to be homologous to peptide A. Synthetic peptide A had a highly inhibitory effect on RADD-integrin α v β 3 binding. The results demonstrate the potential application of short peptides for disrupting high-affinity ADAM-integrin interactions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ADAM:

A Disintegrin And Metalloproteinase

ASMC:

airway smooth muscle cell

b-RADD:

biotinylated RADD

BSA:

bovine serum albumin

DMEM:

Dulbecco modified Eagle medium

EGF:

epidermal growth factor

ELISA:

enzyme-linked immunosorbent assay

GST:

glutathione S-transferase

HBSS:

Hanks balanced salt solution

HMEC:

human microvascular endothelial cells

HPLC:

high-performance liquid chromatography

HRP:

horseradish peroxidase

IL:

interleukin

IPTG:

isopropyl-1-thio-β-D-galactopyranoside

MALDI-TOF:

matrix-assisted laser desorption ionisation time of flight

OPD:

o-phenylenediamine dihydrochloride

PBS:

phosphate buffered saline

RADD:

recombinant human ADAM15 disintegrin domain

RGD:

Arg-Gly-Asp

TBST:

Tris-buffered saline Tween-20

References

  • Arribas J, Bech-Serra J J and Santiago-Josefat B 2006 ADAMs, cell migration and cancer; Cancer Metastasis Rev. 25 57–68

    Article  PubMed  Google Scholar 

  • Azzazy H M and Highsmith W E Jr 2002 Phage display technology: clinical applications and recent innovations; Clin. Biochem. 35 425–445

    Article  PubMed  CAS  Google Scholar 

  • Beck V, Herold H, Benge A, Luber B, Hutzler P, Tschesche H, Kessler H, Schmitt M, Geppert HG, Reuning U 2005 ADAM15 decreases integrin alphavbeta3/vitronectin-mediated ovarian cancer cell adhesion and motility in an RGD-dependent fashion; Int. J. Biochem. Cell Biol. 37 590–603

    Article  PubMed  CAS  Google Scholar 

  • Fleming T J, Sachdeva M, Delic M, Beltzer J, Wescott C R, Devlin M, Lander R C, Nixon A E, Roschke V, Hilbert D M, Sexton D J 2005 Discovery high affinity peptide binders BLyS phage display; J. Mol. Recognit. 18 94–102

    Article  PubMed  CAS  Google Scholar 

  • Jeon O H, Kim D, Choi Y J, Kim S H, Choi W S and Kim D S 2007 Novel function of human ADAM15 disintegrin-like domain and its derivatives in platelet aggregation; Thromb. Res. 119 609–619

    Article  PubMed  CAS  Google Scholar 

  • Karkkainen S, Hiipakka M, Wang J H, Kleino I, Vähä-Jaakkola M, Renkema G H, Liss M, Wagner R, Saksela K 2006 Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome; EMBO Rep. 7 186–191

    Article  PubMed  Google Scholar 

  • Kobayashi N and Yoshida T 2007 Binding sites on laminin receptors as components for antibiotics; Protein Pept. Lett. 14 33–36

    Article  PubMed  CAS  Google Scholar 

  • Kratzschmar J, Lum L and Blobel C P 1996 Metargidin, a membrane-anchored metalloprotease-disintegrin protein with an RGD integrin binding sequence; J. Biol. Chem. 271 4593–4596

    Article  PubMed  CAS  Google Scholar 

  • Lu D, Chung K F, Xia M, Lu X, Scully M and Kakkar V 2006 Integrin binding characteristics of the disintegrin-like domain of ADAM-15; Thromb. Haemost. 96 642–651

    PubMed  CAS  Google Scholar 

  • Lu D, Xie S, Sukkar M B, Lu X, Scully M F and Chung K F 2007 Inhibition of airway smooth muscle adhesion and migration by the disintegrin domain of ADAM-15; Am. J. Respir. Cell Mol. Biol. 37 494–500

    Article  PubMed  CAS  Google Scholar 

  • Mochizuki S and Okada Y 2007 ADAMs in cancer cell proliferation and progression; Cancer Sci. 98 621–628

    Article  PubMed  CAS  Google Scholar 

  • Nath D, Slocombe P M, Stephens P E, Warn A, Hutchinson G R, Yamada K M, Docherty A J and Murphy G 1999 Interaction of metargidin (ADAM-15) with alphavbeta3 and alpha5beta1 integrins on different haemopoietic cells; J. Cell Sci. 112 579–587

    PubMed  CAS  Google Scholar 

  • Steingroewer J, Bley T, Bergemann C and Boschke E 2007 Biomagnetic separation of Salmonella typhimurium with high affine and specific ligand peptides isolated by phage display technique; J. Magn. Magn. Mater. 311 295–299

    Article  CAS  Google Scholar 

  • Stone A L, Kroeger M and Sang Q X 1999 Structure-function analysis of the ADAM family of disintegrin-like and metalloproteinase-containing proteins; J. Protein Chem. 18 447–465

    Article  PubMed  CAS  Google Scholar 

  • Trochon-Joseph V, Martel-Renoir D, Mir L M, Thomaïdis A, Opolon P, Connault E, Li H, Grenet C, Fauvel-Lafève F, Soria J, Legrand C, Soria C, Perricaudet M and Lu H 2004 Evidence of antiangiogenic and antimetastatic activities of the recombinant disintegrin domain of metargidin; Cancer Res. 64 2062–2069

    Article  PubMed  CAS  Google Scholar 

  • Uvebrant K, da Graça Thrige D, Rosén A, Akesson M, Berg H, Walse B and Björk P 2007 Discovery of selective small-molecule CD80 inhibitors; J. Biomol. Screen 12 464–472

    Article  PubMed  CAS  Google Scholar 

  • Wolfsberg T G, Primakoff P, Myles D G and White J M 1995 ADAM, a novel family of membrane proteins containing A Disintegrin And Metalloprotease domain: multipotential functions in cell-cell and cell-matrix interactions; J. Cell Biol. 131 275–278

    Article  PubMed  CAS  Google Scholar 

  • Xiong J P, Stehle T, Zhang R, Joachimiak A, Frech M, Goodman S L and Arnaout M A 2002 Crystal structure of the extracellular segment of integrin alpha Vbeta3 in complex with an Arg-Gly-Asp ligand; Science 296 151–155

    Article  PubMed  CAS  Google Scholar 

  • Zhang X P, Kamata T, Yokoyama K, Puzon-McLaughlin W and Takada Y 1998 Specific interaction of the recombinant disintegrin-like domain of MDC-15 (metargidin, ADAM-15) with integrin alphavbeta3; J. Biol. Chem. 273 7345–7350

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, Jiangsu, 214122, China

    Jing Wu, Min-Chen Wu, Lian-Fen Zhang, Jian-Yong Lei, Lei Feng & Jian Jin

  2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, 214 122, China

    Jing Wu, Min-Chen Wu, Lian-Fen Zhang, Jian-Yong Lei, Lei Feng & Jian Jin

Authors
  1. Jing Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Min-Chen Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lian-Fen Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jian-Yong Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lei Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jian Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jian Jin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, J., Wu, MC., Zhang, LF. et al. Identification of binding peptides of the ADAM15 disintegrin domain using phage display. J Biosci 34, 213–220 (2009). https://doi.org/10.1007/s12038-009-0025-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12038-009-0025-3

Keywords

Search

Navigation