Fluorescein-conjugated lysine monomers for solid phase synthesis of fluorescent peptides and PNA oligomers

TY - JOUR

T1 - Fluorescein-conjugated lysine monomers for solid phase synthesis of fluorescent peptides and PNA oligomers

AU - Lohse, J

AU - Harrit, N

AU - Dahl, O

AU - Nielsen, Peter E.

PY - 1997/7/1

Y1 - 1997/7/1

N2 - Fluorescein ethyl ester, I, was used to prepare the fluorescent mixed ester/ether 6-O-(carboxymethyl)-fluorescein ethyl ester, III. Conjugation of III to the epsilon-amino group of alpha-N-Boc-L-lysine, via the N-hydroxysuccinimde ester, IV, gave the Boc-protected fluorescein-conjugated lysine monomer V. Removal of the Boc group, followed by reaction with Fmoc chloride, gave the Fmoc-protected monomer, VI (Figure 1). These Boc- and Fmoc-protected fluorescein-conjugated lysines were readily incorporated into peptides and PNA oligomers during solid phase synthesis to give fluorescent products. Mass spectroscopy and UV studies showed that the fluorophore remains unchanged during solid phase synthesis. In contrast to fluorescein, the photophysical properties of these derivatives are pH independent from pH 3 to 8, with a molar absorption coefficient, epsilon max 456, of 2.9 x 10(4) M-1 cm-1 and fluorescence quantum yield, phi f, of 0.18.

AB - Fluorescein ethyl ester, I, was used to prepare the fluorescent mixed ester/ether 6-O-(carboxymethyl)-fluorescein ethyl ester, III. Conjugation of III to the epsilon-amino group of alpha-N-Boc-L-lysine, via the N-hydroxysuccinimde ester, IV, gave the Boc-protected fluorescein-conjugated lysine monomer V. Removal of the Boc group, followed by reaction with Fmoc chloride, gave the Fmoc-protected monomer, VI (Figure 1). These Boc- and Fmoc-protected fluorescein-conjugated lysines were readily incorporated into peptides and PNA oligomers during solid phase synthesis to give fluorescent products. Mass spectroscopy and UV studies showed that the fluorophore remains unchanged during solid phase synthesis. In contrast to fluorescein, the photophysical properties of these derivatives are pH independent from pH 3 to 8, with a molar absorption coefficient, epsilon max 456, of 2.9 x 10(4) M-1 cm-1 and fluorescence quantum yield, phi f, of 0.18.

KW - Biopolymers

KW - Chromatography, High Pressure Liquid

KW - Fluorescein

KW - Fluoresceins/chemistry

KW - Lysine/chemistry

KW - Mass Spectrometry/methods

KW - Nucleic Acids/chemical synthesis

KW - Peptides/chemical synthesis

KW - Spectrophotometry, Ultraviolet

U2 - 10.1021/bc9700704

DO - 10.1021/bc9700704

M3 - Journal article

C2 - 9258448

SN - 1043-1802

VL - 8

SP - 503

EP - 509

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 4

ER -