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Identification of Bilirubin Binding Site in Type I Collagen

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Abstract

The interaction of bilirubin with collagen in the significance of jaundice incidence have been previously reported and investigated. The novel peptide sequences containing bilirubin binding domain was identified and located to develop a basis for further studies investigating the interactions of collagen with bilirubin in the present study. In this study an intricate interaction between bilirubin and collagen was characterized and their binding domain has been established using in-gel digestion and LC–MS/MS analysis based on the collagen sequencing and peptide mass fingerprinting. The biotinylated bilirubin derivatives bind to α1(I) chain but not to α2(I) chains which clearly designates that bilirubin shows greater affinity to α1 chains of collagen. The intact proteins collected after analyzing the resulting complex mixture of peptides was used for peptide mapping. Using the electrospray method, among the other peptide sequence information obtained, the molecular weight of collagen alpha-2(I) chain was obtained by locating a 130 kDa weight peptide sequences with greater pi value (9.14) with 1,364 amino acid residues and collagen alpha-1(I) chain with 1,463 amino acid residues with 138.9 kDa molecular weight. This information leads to locate the exact sequence of these helices focussing on the domain identification. The total charge of the peptide domain sequences infers that the bilirubin participates in the electrostatic mode of interaction with collagen peptide. Moreover, other modes of interactions such as hydrogen bonding, covalent interactions and hydrophobic interactions are possible.

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Acknowledgments

The Author (Usharani Nagarajan) was grateful C-CAMP/NCBS MS Facility and Dr. Dominik Schwudke for their support and technical services developed through Chromous Biotech Pvt Ltd.

Conflict of interest

Usharani Nagarajan, Gladstone Christopher Jayakumar, Swarna V Kanth, Jonnalagadda Raghava Rao, Bangaru Chandrasekaran and Balachandran Unni Nair declare that they have no conflict of interest.

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Authors and Affiliations

  1. Centre for Human and Organisational Resources Development, Council of Scientific and Industrial Research (CSIR)-Central Leather Research Institute, Adyar, Chennai, 600020, India

    Nagarajan Usharani, Swarna V. Kanth & Bangaru Chandrasekaran

  2. Chemical Laboratory, Council of Scientific and Industrial Research (CSIR)-Central Leather Research Institute, Adyar, Chennai, 600020, India

    Gladstone Christopher Jayakumar, Jonnalagadda Raghava Rao & Balachandran Unni Nair

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  1. Nagarajan Usharani
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  2. Gladstone Christopher Jayakumar
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  3. Swarna V. Kanth
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  4. Jonnalagadda Raghava Rao
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  5. Bangaru Chandrasekaran
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  6. Balachandran Unni Nair
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Correspondence to Bangaru Chandrasekaran.

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Usharani, N., Jayakumar, G.C., Kanth, S.V. et al. Identification of Bilirubin Binding Site in Type I Collagen. Int J Pept Res Ther 19, 357–364 (2013). https://doi.org/10.1007/s10989-013-9359-7

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  • DOI: https://doi.org/10.1007/s10989-013-9359-7

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