Structures and sites of nonenzymatic glucosylation of protein: characterization by carbon-13 nuclear magnetic resonance spectroscopy

Thesis/Dissertation · Tue Jan 01 04:00:00 EST 1985

OSTI ID:5531645

Fisher, C I

Products of nonenzymatic glucosylation of protein were studied by carbon-13 NMR spectroscopy at 100 MHz in phosphate buffer. The anomeric region of the spectrum of glycated cytochrome c (blocked amino terminus) showed three signals representing the C-2 carbon of the alpha and beta-furanose (101.7 and 99.2 ppm) and beta pyranose (95.8 ppm) anomers of the Amadori rearrangement products (ARP) at lysine epsilon amino groups. The spectrum of glycated poly-L-lysine showed two distinct sets of three resonances; the second set, resonating to lower shielding than the first, was assigned to C-2 of the anomers of ARP at the alpha amino terminus. A third set of peaks was observed on glucation of RNase A corresponding to the ARP at lysine residue 41. The anomeric region of RNase denatured with urea at pH 7.4 was similar to that of glycated poly-L-lysine, while at pH 4.0 it was indistinguishable from that of glycated cytochrome c. The anomeric spectum of glycated hemoglobin was comparable to that of poly-L-lysine, and distinct resonances were observed for the ARP at the alpha amino terminal valine and epsilon amino groups. Because of the differences in chemical shift of C-2 of the ARP, we conclude that the pKa of the amino group influences the electronic environment of the anomeric carbon of the ARP.

• Research Organization: South Carolina Univ., Columbia (USA)
• OSTI ID: 5531645
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
CARBON 13
CARBON ISOTOPES
CARBOXYLIC ACIDS
CYTOCHROMES
EVEN-ODD NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LYSINE
MAGNETIC RESONANCE
METABOLISM
NUCLEAR MAGNETIC RESONANCE
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHATES
PHOSPHORUS COMPOUNDS
PIGMENTS
PROTEINS
RESONANCE
STABLE ISOTOPES
TRACER TECHNIQUES