A designed glycoprotein analogue of Gc-MAF exhibits native-like phagocytic activity

Federica Bogani, Elizabeth McConnell, Lokesh Joshi, Yung Chang, Giovanna Ghirlanda

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Rational protein design has been successfully used to create mimics of natural proteins that retain native activity. In the present work, de novo protein engineering is explored to develop a mini-protein analogue of Gc-MAF, a glycoprotein involved in the immune system activation that has shown anticancer activity in mice. Gc-MAF is derived in vivo from vitamin D binding protein (VDBP) via enzymatic processing of its glycosaccharide to leave a single GalNAc residue located on an exposed loop. We used molecular modeling tools in conjunction with structural analysis to splice the glycosylated loop onto a stable three-helix bundle (α3W, PDB entry 1LQ7). The resulting 69-residue model peptide, MM1, has been successfully synthesized by solid-phase synthesis both in the aglycosylated and the glycosylated (GalNAc-MM1) form. Circular dichroism spectroscopy confirmed the expected α-helical secondary structure. The thermodynamic stability as evaluated from chemical and thermal denaturation is comparable with that of the scaffold protein, α3W, indicating that the insertion of the exogenous loop of Gc-MAF did not significantly perturb the overall structure. GalNAc-MM1 retains the macrophage stimulation activity of natural Gc-MAF; in vitro tests show an identical enhancement of Fc-receptor-mediated phagocytosis in primary macrophages. GalNAc-MM1 provides a framework for the development of mutants with increased activity that could be used in place of Gc-MAF as an immunomodulatory agent in therapy.

Original languageEnglish
Pages (from-to)7142-7143
Number of pages2
JournalJournal of the American Chemical Society
Volume128
Issue number22
DOIs
Publication statusPublished - Jun 7 2006

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Glycoproteins
Proteins
Macrophages
Vitamin D-Binding Protein
Protein Engineering
Solid-Phase Synthesis Techniques
Fc Receptors
Circular dichroism spectroscopy
Circular Dichroism
Thermodynamics
Phagocytosis
Denaturation
Molecular modeling
Immune system
Vitamins
Immune System
Scaffolds (biology)
Spectrum Analysis
Structural analysis
Hot Temperature

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

A designed glycoprotein analogue of Gc-MAF exhibits native-like phagocytic activity. / Bogani, Federica; McConnell, Elizabeth; Joshi, Lokesh; Chang, Yung; Ghirlanda, Giovanna.

In: Journal of the American Chemical Society, Vol. 128, No. 22, 07.06.2006, p. 7142-7143.

Research output: Contribution to journalArticle

Bogani, Federica ; McConnell, Elizabeth ; Joshi, Lokesh ; Chang, Yung ; Ghirlanda, Giovanna. / A designed glycoprotein analogue of Gc-MAF exhibits native-like phagocytic activity. In: Journal of the American Chemical Society. 2006 ; Vol. 128, No. 22. pp. 7142-7143.
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