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Partial alanine scan of mast cell degranulating peptide (MCD): importance of the Histidine‐ and Arginine residues
Author(s) -
Buku Angeliki,
Mendlowitz Milton,
Condie Barry A.,
Price Joseph A.
Publication year - 2004
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.532
Subject(s) - chemistry , alanine , histidine , histamine , peptide , mast cell , fluorescence anisotropy , arginine , binding site , amino acid , receptor , stereochemistry , biochemistry , affinities , pharmacology , biology , immunology , membrane
The influence of the two histidine and two arginine residues of mast cell degranulating peptide (MCD) in activity and binding was studied by replacing these amino acids in the MCD sequence with L ‐alanine. Their histamine releasing activity was determined on rat peritoneal mast cells. Their binding affinity to the FcεRIα binding subunit of the human mast cell receptor protein, was carried out using fluorescence polarization. The histamine assay showed that replacement of His 13 by Ala occurred without loss of activity compared with the activity of MCD. Alanine substitutions for Arg 7 and His 8 resulted in an approximately 40‐fold increase, and for Arg 16 in a 14‐fold increase in histamine‐releasing activity of MCD. The binding affinities of the analogs were tested by competitive displacement of bound fluorescent MCD peptide from the FcεRIα binding protein of the mast cell receptor by the Ala analogs using fluorescence polarization. The analogs Ala 8 (for His) and Ala 16 (for Arg) showed the same binding affinities as MCD, whereas analog Ala 7 (for Arg) and analog Ala 13 (for His) showed slightly better binding affinity than the parent compound. This study showed that the introduction of alanine residues in these positions resulted in MCD agonists of diverse potency. These findings will be useful in further MCD structure–activity studies. Copyright © 2004 European Peptide Society and John Wiley & Sons, Ltd.