Glycosylation enhances functional stability of the chemotactic cytokine CCL2.
The human chemokine CCL2 gene was expressed in the yeast P.pastoris and gave rise to a mixture of differently glycosylated recombinant proteins. In comparison to non-glycosylated E.coli-derived CCL2, glycosylated yeast CCL2L was 4-20 times less active in a chemotactic assay in vitro. However, CCL2L could maintain full activity upon prolonged incubation at 37 degrees C, whereas the non-glycosylated chemokine readily lost activity. It could be hypothesized that glycosylation is a mechanism used by the organism to modulate CCL2 stability. The partial loss of specific activity due to glycosylation is balanced by the advantage of prolonging the effectiveness of chemokine. Thus, differential glycosylation allows one to obtain highly effective short-lived CCL2 or less-effective long-lived CCL2 and may thus represent a novel mechanism of adaptation to pathological versus physiological conditions.
Ruggiero
Paolo
P
Dompé Research Centre, L'Aquila, Italy. borasc@tin.it
Flati
Silvio
S
Di Cioccio
Vito
V
Maurizi
Giovanni
G
Macchia
Giovanni
G
Facchin
Alberto
A
Anacardio
Roberto
R
Maras
Antonio
A
Lucarelli
Marilena
M
Boraschi
Diana
D
D018925:Chemokines / Q000235:genetics / Q000502:physiology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D018925:Chemokines; D018076:DNA, Complementary; D011994:Recombinant Proteins