Structural investigations of the major allergen Phl p I on the complementary DNA and protein level Arnd Petersen, PhD, Gabriele Schramm, MSc, Albrecht.

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Structural investigations of the major allergen Phl p I on the complementary DNA and protein level Arnd Petersen, PhD, Gabriele Schramm, MSc, Albrecht Bufe, MD, Max Schlaak, MD, Wolf-Meinhard Becker, PhD Journal of Allergy and Clinical Immunology Volume 95, Issue 5, Pages 987-994 (May 1995) DOI: 10.1016/S0091-6749(95)70099-4 Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 1 2D immunoblotting of timothy grass pollen extract showing binding reactivity of group I–directed Mab IG 12. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 2 cDNA and deduced amino acid sequence of clone PHLPl.p1. N terminus, determined already by protein sequencing,31 is underlined. Arrow indicates NH2 terminus of mature protein. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 2 cDNA and deduced amino acid sequence of clone PHLPl.p1. N terminus, determined already by protein sequencing,31 is underlined. Arrow indicates NH2 terminus of mature protein. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 2 cDNA and deduced amino acid sequence of clone PHLPl.p1. N terminus, determined already by protein sequencing,31 is underlined. Arrow indicates NH2 terminus of mature protein. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 3 Expression of clone PHLPl.p1 in E. coli SOLR after IPTG treatment. Western blotting of timothy grass pollen extract and SOLR/p1 cell lysates was performed after induction for 0 and 2 hours. I, Protein staining by india ink; P, patients' pooled sera; 1, group I–directed Mab IG 12; 2, group V–directed Mab Bo 1. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 4 Western blotting of timothy grass pollen to study influence of reduction on Phl p I structure. Pollen extract was treated with sample buffers differing in DTT concentrations: 1, no DTT; 2, 0.08 mmol/L; 3, 0.11 mmol/L; 4, 0.20 mmol/L; 5, 0.27 mmol/L; 6, 0.4 mmol/L; 7, 0.8 mmol/L; 8, 1.6 mmol/L; 9, 20 mmol/L; and 10, 40 mmol/L DTT (normally used). Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions

FIG. 5 Comparison of deduced amino acid sequences of timothy grass Phl p I (clone PHLPl.p1), velvet grass Hol l I (clone Hol l I29, and ryegrass Lol p I (clone Lol P1, precursor allergen R727, 28; positions of isoallergenic variants are underlined [position 45, N, D; position 154; A, G; position 187, I, T; position 223, F, V]). Characters below amino acid sequences indicate that position in alignment is perfectly conserved (asterisk), well conserved (s), or different (blank space). Amino acids said to be similar are A, S, and T; D and E; N and Q; R and K; I, L, M, and V; and F, Y, W. Journal of Allergy and Clinical Immunology 1995 95, 987-994DOI: (10.1016/S0091-6749(95)70099-4) Copyright © 1995 Mosby, Inc. Terms and Conditions