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School of Biosciences
   
   
  
 

Marcos Alcocer

Associate Professor, Faculty of Science

Contact

  • workRoom 49E Nutritional Sciences, North Lab Bldg
    Sutton Bonington Campus
    Sutton Bonington
    Leicestershire
    LE12 5RD
    UK
  • work0115 951 6103
  • fax0115 951 6122

Expertise Summary

My research interests are predominantly: food allergy and protein expression.

Our main expertise are on:

-Food allergens (analysis and production)-Mechanisms of allergenicity-Recombinant protein expression (mainly eukaryote)-Protein structure analysis

Research Summary

My research interests are predominantly in two interrelated areas of research: food allergens and protein expression.

  • In the area of food allergens I am interested in understanding why specific proteins are recognised by the adaptive immune system as "foreign" whilst other protein with similar structures are tolerated. Our working molecular model is based around the 2S albumin/LTP/amylase inhibitor family of food allergenic proteins. We have shown that Ber e 1, the major 2S albumin allergen from brazil nut (Bertoleta excelsa), SFA8, the 2S albumin from sunflower seed, and two 2S albumins from soybean (9) are secreted at high levels by the methylotrophic yeast Pichia pastoris and that the recombinant proteins mimic their native counterparts in structure and biophysical characteristics (1,2). In subsequent studies we demonstrated that those proteins were highly resistant to thermal and chemical-induced de-naturation and proteolytic digestion, important attributes in the assessment of intrinsic allergenicity of proteins (3,4). Our results have suggested Ber e 1 to be the only intact brazil nut seed protein that withstands peptic digestion, making it a prime candidate for recognition by the gut immune system. The recombinant Ber e 1 has been double-labelled (13C/15N) and the final 3D structure determined by NMR is near completion(6). Through the use of a chimaeric protein microarray system, recently developed in our lab, we have mapped the structural IgE epitope of Ber e 1 to a particular helix-loop-helix region (5). The same structural region has been previously reported as the immunodominant region in related food allergens by different techniques. The chimaeric proteins and other recombinant allergens are currently been used to investigate the signals involved during dendritic cell maturation at cellular level (7,8).
  • In the protein expression area, in collaborative work with Prof Archer from the School of Biology, Nottingham and with University of Cambridge, we are interested in understanding how eukaryotic cells (mainly fungi) process, fold and regulate protein secretion. Our working molecular model is based around the human lysozyme protein. Several mutants have now been produced, labelled and analysed at structural level (10,11). There seem to be a good correlation between their tendency to form amyloid aggregates and their low expression in fungal systems. Strategies to understand the signals involved and ways to re-establish the wild type secretion are being developed.

Selected Publications:

1 ALCOCER, M.J.C., MURTAGH, G.J., BAILEY, K., DUMOULIN, M., MESEGUER, A.S., PARKER, M.J. & ARCHER D.B. (2002). The disulphide mapping, folding and chracterisation of recombinant Ber e 1, an allergenic protein and SFA8, two sulphur-rich 2S plant albumins. J. Mol. Biol. 324 (1):165-175.2 LIN, J., FIDO, R., SHEWRY, P., ARCHER, D.B., ALCOCER, M.J.C. (2004) The expression and processing of two recombinant 2S albumins from soybean (Glycine max) in the yeast Pichia pastoris. Biophys. Biochem. Acta, 1698: 203-212. 3 MURTAGH, G.J., ARCHER, D.B., DUMOULIN, M., ALCOCER, M.J.C. (2003) In vitro stability and immunoreactivity of the native and recombinant plant food 2S albumins Ber e 1 and SFA-8. Clin. Exp. Allergy 33:1147-1152.4 MURTAGH, G.J., ARCHER, D.B., DUMOULIN, M., ALCOCER, M.J.C. (2002) Stability of recombinant 2S albumin plant food allergens in vitro. Biochem. Soc. Trans. 30 (6): 913-915. 5 ALCOCER,M.J.C., MURTAGH, G.J., WILSON, P.B., PROGIAS, P., LIN, J., ARCHER, D.B. (2004) The Major Human Structural IgE Epitope of the Brazil Nut Allergen Ber e 1: A Chimaeric and Protein Microarray Approach. J. Mol. Biol. 343:759-7696 TENGEL, T., ALCOCER, M.J.C., SCHLEUCHER, J., LARSSON, G. (2005) Complete assignement and secondary structure of the brazil nut allergen Ber e 1. J. Biomol. NMR, 32(4):3367 KEAN, D.E., GOODRIDGE, H.S., HARNETT, M.M., ALCOCER, M.J.C., HARNETT, W. (2006) Differential polarization of dendritic cells and helper T cell responses by the plant 2S seed albumins Ber e 1 and SFA8. J. Immunology (in print).8 DEARMAN, R., ALCOCER, M.J.C., KIMBER, I. (2006). Influence of plant lipids on immune responses in mice to the major Brazil nut allergen Ber e 1. Clin. Exp. Allergy (submitted)9 LIN, J., SHEWRY, P., ARCHER, D.B., BEYER, K., NIGGEMANN, B., HAAS, H., WILSON, P. ALCOCER, M.J.C. (2006) The potential allergenicity of two 2S albumin from soybean (Glycine max): a protein microarray approach. Int. Arch. Allergy Immunol., (in print)10 KUMITA, J.R., RUSSEL, J.K., ALCOCER, M.J.C., DUMOULIN, M., McCAMMON, M.G., ROBINSON, C.V., ARCHER, D.B., DOBSON, C.M. (2006) Secreted levels and stability of two novel human lysozyme variants (W64R and F57I) associated with hereditary renal amyloidosis. FEBS J., 273: 711-720.11 JOHNSON, R.J.K., CHRISTODOULOU, J., DUMOULIN, M., CADDY, G.L., ALCOCER, M.J.C., MURTAGH, G.J., KUMITA, J.R., LARSSON, G. ROBINSON, C.V., ARCHER, D.B., LUISI, B., DOBSON, C.M. (2005) Rationalising lysozyme amyloidosis:insight from the structure and solution dynamics of T70N lysozyme.J. Mol. Biol., 352(4):823-836.

Selected Publications

Past Research

The ability to design, label and analyse the 3D structure of proteins has generated a series of parallel work in the expression area (1-7). In collaboration with the School of Biology, Nottingham and University of Cambridge work is been carried out in order to understand how eukaryotic cells (mainly fungi) process, fold and regulate protein secretion (3-7). Several mutant proteins have now been produced, labelled and analysed at structural level (1,2). In collaboration with the University of Strathclyde the immunogenicity of the parasitic ES-62 was assessed (3).

Selected publications

1- KUMITA et al., (2006) Secreted levels and stability of two novel human lysozyme variants (W64R and F57I) associated with hereditary renal amyloidosis. FEBS Journal, 273: 711-720.2- JOHNSON et al., (2005) Rationalising lysozyme amyloidosis:insight from the structure and solution dynamics of T70N lysozyme. Journal of Molecular Biology, 352(4):823-836.3- AL-SHEIKH et al., (2004) - Endoplasmic reticulum stress leads to the selective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger. MolecularMicrobiology 53(6):1731-1742.4- DYER et al., (2003). Identification of a mating-type gene in the homothallic fungus Aspergillus nidulans. Fungal Genetics Newsletter 50 (Supplement): 1455- ALCOCER et al., (2003). Comparison of modular and non-modular xylanases as carrier proteins for the efficient secretion of heterologous protein in Penicillium funiculosum. Applied Microbiology and Biotechnology, 60:726-732.6- BELSHAW eta al., (2002). Use of a histone H4 promoter to drive the expression of homologous and heterologous proteins by Penicillium funiculosum. Applied Microb. Biotechnol. 60:455-460.7- FURNISS et al., (2002) A family 11 xylanase from Penicillium funiculosum Is strongly inhibited by three wheat xylanase inhibitors. Biophysical Biochemica Acta. 1598:24-29.8- EGAN et al., (2005) Lack of immunological cross-reactivity between native and recombinant forms of ES-62, a secreted protein of Acanthocheilonema vitae. Parasitology, 132:1-12.

Future Research

The tools that have been generated from previous project have allowed us to embark on large-scale projects involving population screening. For this we are finalising a protein microarray that will contain most of the proteins from the British diet. The use of protein microarrays in association with basophils will become a powerful tool for the understanding of molecular events leading to food allergy.

School of Biosciences

University of Nottingham
Sutton Bonington Campus
Loughborough, Leicestershire LE12 5RD

telephone: +44 (0)115 9516400
fax: +44 (0) 115 951 6020
email: biosciences-enquiries@nottingham.ac.uk