Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides

Yang Li, Daniel Ho, Huan Meng, Tania R. Chan, Bo An, Hanry Yu, Barbara Brodsky, Albert Jun, S. Michael Yu

Research output: Contribution to journalArticle

Abstract

Although fibrous collagens are major structural components of extracellular matrix in mammals, collagen overproduction is associated with many human diseases including cancers and fibrosis. Collagen is typically identified in biomedical research by Western blot and immunohistochemistry; however, anticollagen antibodies employed in these analyses are difficult to prepare and their affinities to collagen can diminish if collagen becomes denatured during analyses. Previously, we discovered that single-stranded collagen mimetic peptides [CMPs, sequence: (GlyProHyp)9] can bind to denatured collagen chains by triple helix hybridization. Here, we present collagen-specific staining methods using simple CMPs conjugated to common fluorophores (e.g., carboxyfluorescein), which allow direct detection of collagens and collagen-like proteins in SDS-PAGE and in various mammalian tissue sections. By directly staining SDS-PAGE gels with fluorescently labeled CMPs, both intact (type I, II, and IV) and MMP-1 cleaved collagen (type I) chains as well as complement factor C1q were detected. Collagen bands containing as little as 5 ng were optically visualized, while no staining was observed for fibronectin, laminin, and a collection of proteins from mammalian cell lysate. The CMP was unable to stain collagen-like bacterial protein, which contains numerous charged amino acids that are believed to stabilize triple helix in place of Hyp. We also show that fluorescently labeled CMPs can specifically visualize collagens in fixed tissue sections (e.g., skin, cornea, and bone) more effectively than anticollagen I antibody, and allow facile identification of pathologic conditions in fibrotic liver tissues.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalBioconjugate Chemistry
Volume24
Issue number1
DOIs
StatePublished - Jan 16 2013

Fingerprint

Collagen
Peptides
Proteins
Cytidine Monophosphate
Tissue
Staining and Labeling
Antibodies
Polyacrylamide Gel Electrophoresis
Complement C1q
Mammals
Bacterial Proteins
Fluorophores
Laminin
Collagen Type I
Matrix Metalloproteinases
Fibronectins
Liver
Cornea
Extracellular Matrix
Biomedical Research

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Li, Y., Ho, D., Meng, H., Chan, T. R., An, B., Yu, H., ... Michael Yu, S. (2013). Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides. Bioconjugate Chemistry, 24(1), 9-16. https://doi.org/10.1021/bc3005842

Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides. / Li, Yang; Ho, Daniel; Meng, Huan; Chan, Tania R.; An, Bo; Yu, Hanry; Brodsky, Barbara; Jun, Albert; Michael Yu, S.

In: Bioconjugate Chemistry, Vol. 24, No. 1, 16.01.2013, p. 9-16.

Research output: Contribution to journalArticle

Li, Y, Ho, D, Meng, H, Chan, TR, An, B, Yu, H, Brodsky, B, Jun, A & Michael Yu, S 2013, 'Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides', Bioconjugate Chemistry, vol. 24, no. 1, pp. 9-16. https://doi.org/10.1021/bc3005842
Li, Yang ; Ho, Daniel ; Meng, Huan ; Chan, Tania R. ; An, Bo ; Yu, Hanry ; Brodsky, Barbara ; Jun, Albert ; Michael Yu, S. / Direct detection of collagenous proteins by fluorescently labeled collagen mimetic peptides. In: Bioconjugate Chemistry. 2013 ; Vol. 24, No. 1. pp. 9-16.
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