Synthesis and characterization of conjugates formed between periodate-oxidized ribonucleotides and amine-containing fluorophores.
The synthesis and purification of new fluorescently labeled derivatives of GDP and ATP are described. The fluorescent groups are coupled initially through amine-containing linker arms to periodate-oxidized nucleotides. Reduction of the initial product yields primarily a six-membered morpholine-like ring. Fluorescein-labeled GDP, rhodamine-labeled GDP, and fluorescein-labeled ATP were characterized by absorbance spectroscopy and TLC. NMR and FAB-MS studies were carried out on a single nucleotide derivative formed by reacting periodate-oxidized guanosine and benzylamine with subsequent reduction to establish the modification to the ribose moiety. The synthesis of the guanosine-benzylamine conjugate led to a mixture of products that were separated. The predominant product (70%) resulted in conversion of the ribose moiety to a six-membered morpholine-like ring having no hydroxyl group, and the minor product (30%) resulted in an open ring structure having one hydroxyl. When NaCNBH3 was used as the sole reductant, only the product with the morpholine-like ring was formed. These probes were prepared for use in solution studies of the interactions of eukaryotic initiation factor-2 with other components of mammalian protein synthesis initiation.
Department of Chemistry, University of Nebraska--Lincoln 68588-0304.
R E
RE
Hileman
K M
KM
Parkhurst
N K
NK
Gupta
L J
LJ
Parkhurst
D000255:Adenosine Triphosphate / Q000737:chemistry; D000588:Amines / Q000737:chemistry*; D015852:Eukaryotic Initiation Factor-2 / Q000737:chemistry; D005452:Fluoresceins / Q000138:chemical synthesis / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006151:Guanosine / Q000737:chemistry; D006153:Guanosine Diphosphate / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry*; D010758:Phosphorus / Q000032:analysis; D012265:Ribonucleotides / Q000737:chemistry*; D012266:Ribose / Q000737:chemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013056:Spectrophotometry, Ultraviolet
D000588:Amines; D015852:Eukaryotic Initiation Factor-2; D005452:Fluoresceins; D005456:Fluorescent Dyes; D012265:Ribonucleotides; D010504:Periodic Acid; D006151:Guanosine; D006153:Guanosine Diphosphate; D010758:Phosphorus; D012266:Ribose; D000255:Adenosine Triphosphate