Selective fluorogenic derivatization of Pro2—Lys peptides with naphthalene-2,3-dicarboxaldehyde/cyanide

✍ Scribed by F. Kristjansson; A. Thakur; J.F. Stobaugh

• Book ID: 102982458
• Publisher: Elsevier Science
• Year: 1992
• Tongue: English
• Weight: 682 KB
• Volume: 262
• Category: Article
• ISSN: 0003-2670
• DOI: 10.1016/0003-2670(92)80057-e

✦ Synopsis

The potential for fluorescence quenclnng due to bls-derwatlzatlon of Lys peptldes with naphthalene-2,3-dlcarboxaldehyde/cyamde was confirmed Kmetlc mvestlgatlon revealed that derwatlzatlon at the c-ammo ates proceeded slgmficantly faster than at a-ammo sites After charactemafion of pH and co-solvent (methanol) effects on the absolute and relatrve reactlon rate at l -and cr-amme srtes, It was concluded that by usmg a combmatlon of pH control and tmung that the formation of the analytlcally useful mono-dernratnre at the E-ammo site was feasible Using this lonetic mformatlon, denvafizatlon at pH 10 with 80% methanol as co-solvent for 2 nun resulted m the formatlon of a htghly fluorescent mono-denvatme of the Pro'-Lys peptIde, Substance P When tins derrvatrzatlon protocol was used m conpmctlon with hqmd chromatography and fluorescence detection for Substance P, a hnear cahbratlon plot resulted (SO-500 nM range, 194-19 4 pmol mjected, R S D 5 2% at each pomt)