Human recombinant alpha 2-HS glycoprotein is produced in insect cells as a full length inhibitor of the insulin receptor tyrosine kinase.
Human Alpha 2-HS glycoprotein (AHSG), a glycoprotein synthesized by hepatocytes, was expressed in insect cells using the recombinant baculovirus system. The protein was purified from the cell supernatant, and appeared as a single band at about 52 kDa. Western blot using a specific antibody to the B-chain of AHSG indicated that the connecting peptide was present in the protein. When incubated with solubilized insulin receptors, recombinant AHSG inhibited the tyrosine kinase activity of the receptors in a dose-dependent fashion at concentrations in the range of those of the circulating protein. AHSG did not interfere with the binding of insulin to its receptor. These results indicate that human AHSG represents a natural inhibitor of the insulin receptor tyrosine kinase, is active as a single-chain protein and possesses a biological role similar to that of its homologue in rats, pp63, described by Auberger et al. (1).
Kalabay
L
L
3rd Department of Medicine, Semmelweis University Medical School, Budapest, Hungary.
Chavin
K
K
Lebreton
J P
JP
Robinson
K A
KA
Buse
M G
MG
Arnaud
P
P
D016367:Baculoviridae / Q000235:genetics; D001798:Blood Proteins / Q000494:pharmacology*; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability / Q000502:physiology; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology; D008099:Liver / Q000502:physiology; D010766:Phosphorylation; D011485:Protein Binding; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis / Q000037:antagonists & inhibitors*; D011972:Receptor, Insulin / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D060749:alpha-2-HS-Glycoprotein
C556354:AHSG protein, human; D001798:Blood Proteins; D004791:Enzyme Inhibitors; D007328:Insulin; D011994:Recombinant Proteins; D060749:alpha-2-HS-Glycoprotein; D020794:Receptor Protein-Tyrosine Kinases; D011972:Receptor, Insulin