Lysine vasopressin undergoes rapid glycation in the presence of reducing sugars.
Lysine vasopressin (LVP) readily reacts with reducing saccharides both in lyophilized preparations and in aqueous solution. Incubation of LVP with, for example, lactose over a pH range of 3.0-8.5 in phosphate buffer or simply in water, gives rise to a number of reaction products, some of which form rapidly (in a matter of hours) even in the frozen state. Reaction mixtures were analysed by reversed-phase HPLC and the structures of the products were deduced from the amino-acid composition of isolated components, by comparison with product profiles obtained with analogues under similar conditions and by FAB mass-spectral analysis of derivatives isolated after reduction with cyanoborohydride. The primary products arise from the formation of Schiff's bases at one or both of the two amino functions. The alpha-amino group of the N-terminal cystine is considerably more reactive than is the epsilon-amino group of lysine and it is the N-terminal adduct which rapidly forms even at -20 degrees C. It is concluded that caution must be shown in using reducing sugars in formulations containing peptides and proteins, particularly the vasopressins and oxytocin.
Division of Chemistry, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, UK.
E
E
Tarelli
P H
PH
Corran
B R
BR
Bingham
H
H
Mollison
R
R
Wait
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D005612:Freeze Drying; D006031:Glycosylation; D007785:Lactose / Q000032:analysis / Q000737:chemistry*; D008236:Lypressin / Q000737:chemistry*; D008969:Molecular Sequence Data; D013696:Temperature
D008236:Lypressin; D007785:Lactose