Effect of carbohydrate structure on biological activity of artificially N-glycosylated eel calcitonin.
To reveal the function of the carbohydrate portion of glycopeptides and glycoproteins, we chemo-enzymatically synthesized artificially N-glycosylated derivatives of eel calcitonin and studied their three-dimensional structure and biological activity. The CD and NMR spectra in trifluoroethanol-H(2)O solution showed that the glycosylation did not change the three-dimensional structure. All the derivatives retained the strong in vivo hypocalcemic activity of calcitonin. However, the relative activity was dependent on the structure of the attached carbohydrate. The single GlcNAc attachment best enhanced the activity, while larger carbohydrates decreased the activity. This relative activity order of compounds could be partly explained by their calcitonin-receptor binding affinity, though the affinity of the GlcNAc derivative did not exceed that of calcitonin. The enhanced hypocalcemic activity of the GlcNAc derivative was explained by its altered biodistribution. The GlcNAc attachment caused calcitonin to escape from the trap at the liver during the early circulation. Thus, the glycosylation was shown to modulate the biological activity of calcitonin depending on the carbohydrate structure without a change in the peptide backbone conformation.
Tagashira
M
M
Analytical Research Laboratory, Asahi Kasei Corporation, Fuji, Shizuoka 416-8501, Japan.
Tanaka
A
A
Hisatani
K
K
Isogai
Y
Y
Hori
M
M
Takamatsu
S
S
Fujibayashi
Y
Y
Yamamoto
K
K
Haneda
K
K
Inazu
T
T
Toma
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002116:Calcitonin / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis* / Q000737:chemistry*; D002942:Circular Dichroism; D004524:Eels / Q000378:metabolism*; D006031:Glycosylation; D006996:Hypocalcemia / Q000139:chemically induced; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D011487:Protein Conformation; D018003:Receptors, Calcitonin / Q000378:metabolism; D013329:Structure-Activity Relationship; D013696:Temperature; D013997:Time Factors; D014018:Tissue Distribution
D002241:Carbohydrates; D018003:Receptors, Calcitonin; D002116:Calcitonin