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Tryptophan residue is essential for immunoreactivity of a diagnostically relevant peptide epitope of A. fumigatus

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Abstract

The role of tryptophan (Trp17) in immunoreactivity of P1, the diagnostically relevant peptide from a major allergen/antigen of Aspergillus fumigatus, was evaluated by chemically modifying tryptophanyl residue of P1. In BIAcore kinetic studies, unmodified P1 showed a 100-fold higher binding with ABPA (Allergic Bronchopulmonary Aspergillosis) patients’ IgG [KD (equilibrium dissociation constant) = 2.74 e−8 ± 0.13 M] than the controls’ IgG (KD = 2.97 e−6± 0.14 M), whereas chemically-modified P1 showed similar binding [KD patients’ IgG = 3.25 e−7± 0.16 M, KD controls’ IgG = 3.86 e−7± 0.19 M] indicating loss of specific immunoreactivity of P1 on tryptophan modification. Modified P1 showed loss of specific binding to IgE and IgG antibodies of ABPA patients in ELISA (Enzyme-Linked Immunosorbent Assay). The study infers that tryptophan residue (Trp17) is essential for immunoreactivity of P1.

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Abbreviations

Afu :

Aspergillus fumigatus

ABPA:

Allergic bronchopulmonary aspergillosis

Asp f 1 :

An allergen/antigen/ cytotoxin of A. fumigatus

ELISA:

Enzyme-linked immunosorbent assay

TFE:

Tri fluoro ethanol

NBS:

N-bromosuccinimide

K D :

Equilibrium dissociation constant

k d :

Dissociation rate constant

k a :

Association rate constant

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Author notes

  1. Deepak Sharma

    Present address: Biology Department, Boston College, Chestnut Hill, Massachusetts, U.S.A.

  2. P. Usha Sarma

    Present address: Department of Plant Pathology, Indian Agricultural Research Institute, Delhi, India

Authors and Affiliations

  1. Institute of Genomics and Integrative Biology, Mall Road, Delhi, India

    Neel Kamal, Shantanu Chowdhury, Taruna Madan & P. Usha Sarma

  2. Department of Chemistry, University of Delhi, Delhi, India

    Deepak Sharma & M. Attreyi

  3. Biopolymers Division, Central Drug Research Institute, Lucknow, India

    Wahajul Haq & Seturam Bandacharya Katti

  4. School of Biotechnology, Devi Ahilya Vishwavidyalaya, Khandwa Road, Indore, India

    Neel Kamal & Anil Kumar

  5. Molecular Biochemistry and Diagnostics Division, Institute of Genomics and Integrative Biology, Mall Road, Delhi-7, India

    Taruna Madan

Authors
  1. Neel Kamal
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  2. Shantanu Chowdhury
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Correspondence to Taruna Madan.

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Kamal, N., Chowdhury, S., Madan, T. et al. Tryptophan residue is essential for immunoreactivity of a diagnostically relevant peptide epitope of A. fumigatus. Mol Cell Biochem 275, 223–231 (2005). https://doi.org/10.1007/s11010-005-2056-x

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  • DOI: https://doi.org/10.1007/s11010-005-2056-x

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