Abstract
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 FcERIα binding subunit of the human mast cell receptor protein, was carried out using fluorescence polarization. The histamine assay showed that replacement of His13 by Ala occurred without loss of activity compared with the activity of MCD. Alanine substitutions for Arg7 and His8 resulted in an approximately 40-fold increase, and for Arg16 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 FcERIα binding protein of the mast cell receptor by the Ala analogs using fluorescence polarization. The analogs Ala8 (for His) and Ala16 (for Arg) showed the same binding affinities as MCD, whereas analog Ala7 (for Arg) and analog Ala13 (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.
| Original language | English |
|---|---|
| Pages (from-to) | 313-317 |
| Number of pages | 5 |
| Journal | Journal of Peptide Science |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| State | Published - 1 Jun 2004 |
Fingerprint
Keywords
- Ala-scan
- Flourescence polarization
- Histamine release
- MCD
- Mast cell receptor
Cite this
}
Partial alanine scan of mast cell degranulating peptide (MCD) : Importance of the histidine- and arinine residues. / Buku, Angeliki; Mendlowitz, Milton; Condie, Barry A.; Price, Joseph.
In: Journal of Peptide Science, Vol. 10, No. 6, 01.06.2004, p. 313-317.Research output: Contribution to journal › Article
TY - JOUR
T1 - Partial alanine scan of mast cell degranulating peptide (MCD)
T2 - Importance of the histidine- and arinine residues
AU - Buku, Angeliki
AU - Mendlowitz, Milton
AU - Condie, Barry A.
AU - Price, Joseph
PY - 2004/6/1
Y1 - 2004/6/1
N2 - 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 FcERIα binding subunit of the human mast cell receptor protein, was carried out using fluorescence polarization. The histamine assay showed that replacement of His13 by Ala occurred without loss of activity compared with the activity of MCD. Alanine substitutions for Arg7 and His8 resulted in an approximately 40-fold increase, and for Arg16 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 FcERIα binding protein of the mast cell receptor by the Ala analogs using fluorescence polarization. The analogs Ala8 (for His) and Ala16 (for Arg) showed the same binding affinities as MCD, whereas analog Ala7 (for Arg) and analog Ala13 (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.
AB - 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 FcERIα binding subunit of the human mast cell receptor protein, was carried out using fluorescence polarization. The histamine assay showed that replacement of His13 by Ala occurred without loss of activity compared with the activity of MCD. Alanine substitutions for Arg7 and His8 resulted in an approximately 40-fold increase, and for Arg16 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 FcERIα binding protein of the mast cell receptor by the Ala analogs using fluorescence polarization. The analogs Ala8 (for His) and Ala16 (for Arg) showed the same binding affinities as MCD, whereas analog Ala7 (for Arg) and analog Ala13 (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.
KW - Ala-scan
KW - Flourescence polarization
KW - Histamine release
KW - MCD
KW - Mast cell receptor
UR - http://www.scopus.com/inward/record.url?scp=3042665907&partnerID=8YFLogxK
U2 - 10.1002/psc.532
DO - 10.1002/psc.532
M3 - Article
C2 - 15214434
AN - SCOPUS:3042665907
VL - 10
SP - 313
EP - 317
JO - Journal of Peptide Science
JF - Journal of Peptide Science
SN - 1075-2617
IS - 6
ER -