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Synthesis and molecular characterization of a biotinylated analog of [Lys]bradykinin
- Source :
- Peptides. 12:1019-1024
- Publication Year :
- 1991
- Publisher :
- Elsevier BV, 1991.
-
Abstract
- We report the synthesis and molecular characterization of a biotinylated analog of kallidin, [Lys]bradykinin. Bradykinin was prepared by solid phase peptide synthesis. Before cleavage from the resin, a biotin moiety was coupled to the epsilon amino group of a lysine in the zeroth position of the bradykinin peptide. An omega-amino caproic acid spacer was incorporated between the biotin group and the N-terminal lysine. The biotinylated peptide was deprotected, cleaved from the resin and purified by RP-HPLC. The identity of this analog was confirmed by amino acid analysis and FAB-mass spectrometry. Biotinyl [Lys]bradykinin (BLBK, mol, wt. = 1528) inhibited [3H]-bradykinin binding to guinea pig ileum homogenates dose dependently, with an IC50 of 28.9 +/- 6 nM. The IC50 for [Lys]bradykinin was approximately 10-fold lower, 3.2 +/- 0.6 nM. BLBK induced contractility in an isolated guinea pig smooth muscle preparation with an EC50 of 129 +/- 14 nM; the corresponding value for [Lys]bradykinin was 29 +/- 8 nM. These data are consistent with the difference in binding potency observed for BLBK compared to [Lys]bradykinin. In an ELISA assay using BLBK and affinity-purified rabbit anti-bradykinin antibody, BLBK bound to anti-bradykinin antibody with an EC50 = 1.21 +/- 0.54 nM. Rank order potencies for several bradykinin peptide analogs suggest that the epitope on bradykinin recognized by the antibody is likely to be at the carboxy terminus of the peptide.
- Subjects :
- Physiology
Stereochemistry
Guinea Pigs
Molecular Sequence Data
Lysine
Bradykinin
Peptide
In Vitro Techniques
Biochemistry
Structure-Activity Relationship
Cellular and Molecular Neuroscience
chemistry.chemical_compound
Endocrinology
Biotin
Ileum
Peptide synthesis
Animals
Amino Acid Sequence
Binding site
chemistry.chemical_classification
Dose-Response Relationship, Drug
Molecular Structure
Kallidin
Receptors, Bradykinin
Cell Membrane
Muscle, Smooth
Receptors, Neurotransmitter
Kinetics
chemistry
Biotinylation
Indicators and Reagents
Muscle Contraction
Subjects
Details
- ISSN :
- 01969781
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Peptides
- Accession number :
- edsair.doi.dedup.....84e8e0b20b382ded035da0736329e503