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Modeling the human intestinal Mucin (MUC2) C-terminal cystine knot dimer

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Abstract

Intestinal mucus, a viscous secretion that lines the mucosa, is believed to be a barrier to absorption of many therapeutic compounds and carriers, and is known to play an important physiological role in controlling pathogen invasion. Nevertheless, there is as yet no clear understanding of the barrier properties of mucus, such as the nature of the molecular interactions between drug molecules and mucus components as well as those that govern gel formation. Secretory mucins, large and complex glycoprotein molecules, are the principal determinants of the viscoelastic properties of intestinal mucus. Despite the important role that mucins play in controlling transport and in diseases such as cystic fibrosis, their structures remain poorly characterized. The major intestinal secretory mucin gene, MUC2, has been identified and fully sequenced. The present study was undertaken to determine a detailed structure of the cysteine-rich region within the C-terminal end of human intestinal mucin (MUC2) via homology modeling, and explore possible configurations of a dimer of this cysteine-rich region, which may play an important role in governing mucus gel formation. Based on sequence–structure alignments and three-dimensional modeling, a cystine knot tertiary structure homologous to that of human chorionic gonadotropin (HCG) is predicted at the C-terminus of MUC2. Dimers of this C-terminal cystine knot (CTCK) were modeled using sequence alignment based on HCG and TGF-beta, followed by molecular dynamics and simulated annealing. Results support the formation of a cystine knot dimer with a structure analogous to that of HCG.

Homodimer model of human intestinal mucin (MUC2) in ribbon presentation with the chains colored red and blue. Human chorionic gonadotropin (HCG) CysX model based on the 3-D structure of (1HCN) with an inter-chain disulfide bond between Cys54 of each chain

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Authors and Affiliations

  1. Department of Chemical Engineering, Northeastern University, 342 Snell Engineering Center, 360 Huntington Avenue, Boston, MA, 02115, USA

    Vatsala D. Sadasivan, Albert Sacco Jr & Rebecca L. Carrier

  2. Department of Biology, Northeastern University, 134 Mugar Life Sciences Building, 360 Huntington Avenue, Boston, MA, 02115, USA

    Sandeep R. Narpala

  3. Department of Chemistry and Chemical Biology, Northeastern University, 102 Hurtig Hall, 360 Huntington Avenue, Boston, MA, 02115, USA

    David E. Budil

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  1. Vatsala D. Sadasivan
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Correspondence to Rebecca L. Carrier.

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Supplementary Fig. 1

ANOLEA [58] method assessing the atomic mean force potential. Negative energy values (green) represent a favorable environment. whereas positive values (red) represent an unfavorable environment for each amino acid. a 3D model of the C-terminus of MUC2, b HCG-B (GIF 27 kb)

High resolution image file (TIFF 27171 kb)

Supplementary Fig. 2

Ramachandran plots generated by the PROCHECK [59] program displaying psi (ψ) and phi (φ) dihedral angles of each residue in the protein structure to assess the “stereochemical quality” of a structure. Red Conformations of ψ and φ that have no steric clashes; white sterically disallowed for all amino acids except glycine (represented as triangles in both plots) a 3D model of the C-terminus of MUC2, b HCG-B (GIF 39 kb)

High resolution image file (TIFF 49024 kb)

Supplementary Fig. 3

Ribbon presentation of the two dimer models based on the 3D structure of human chorionic gonadotropin (PDB ID: 1HCN). HCG CysX is colored yellow and HCG noCysX is colored blue (GIF 65 kb)

High resolution image file (TIFF 13584 kb)

Supplementary Fig. 4

All three MUC2 dimer models are represented in Corey-Pauling-Koltun (CPK) representation. a HCG CysX, b HCG noCysX, c TGFβ. Hydrophobic residues are colored grey and all other residues are colored green (GIF 383 kb)

High resolution image file (TIFF 73063 kb)

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Sadasivan, V.D., Narpala, S.R., Budil, D.E. et al. Modeling the human intestinal Mucin (MUC2) C-terminal cystine knot dimer. J Mol Model 17, 2953–2963 (2011). https://doi.org/10.1007/s00894-010-0932-0

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