Labelling of peptides with 1.4-nm gold particles to demonstrate their binding sites in the rat spinal cord.
Recently we presented a method to label the neuropeptide substance P with a 1.4-nm gold particle covalently bound at the N-terminus that can be used for demonstrating its binding sites in histological sections. In this study we examined whether the peptides neuropeptide Y, somatostatin, calcitonin gene-related peptide and bradykinin can be labelled in the same way. Polyacrylamide gel electrophoresis revealed a reduction in mobility for peptide-gold conjugates over gold particles alone consistent with peptide binding. In cryostat sections of the rat lumbar spinal cord, the peptides showed a distinct binding pattern in the grey matter corresponding to data of studies using autoradiographic methods. Therefore, we conclude that this simple and fast method can be used for labelling peptides in general to demonstrate their binding sites in histological sections, provided the peptide binds by its C-terminus.
Physiologisches Institut der Universität Würzburg, Röntgenring 9, D-97070 Würzburg, Germany.
K
K
Schmidt
G
G
Segond von Banchet
B
B
Heppelmann
D000818:Animals; D001665:Binding Sites / Q000187:drug effects / Q000502:physiology; D001920:Bradykinin / Q000032:analysis / Q000493:pharmacokinetics; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis / Q000493:pharmacokinetics; D017979:Gold Colloid / Q000737:chemistry* / Q000493:pharmacokinetics*; D008297:Male; D009474:Neurons / Q000737:chemistry* / Q000166:cytology / Q000187:drug effects; D009478:Neuropeptide Y / Q000032:analysis / Q000493:pharmacokinetics; D009479:Neuropeptides / Q000032:analysis* / Q000493:pharmacokinetics*; D020671:Posterior Horn Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D013004:Somatostatin / Q000032:analysis / Q000493:pharmacokinetics; D013116:Spinal Cord / Q000737:chemistry* / Q000166:cytology / Q000187:drug effects
D017979:Gold Colloid; D009478:Neuropeptide Y; D009479:Neuropeptides; D013004:Somatostatin; D015740:Calcitonin Gene-Related Peptide; D001920:Bradykinin