Constitutive secretion of serum albumin requires reversible protein tyrosine phosphorylation events in trans-Golgi.
Serum albumin secretion from rat hepatocytes proceeds via the constitutive pathway. Although much is known about the role of protein tyrosine phosphorylation in regulated secretion, nothing is known about its function in the constitutive process. Here we show that albumin secretion is inhibited by the tyrosine kinase inhibitor genistein but relatively insensitive to subtype-selective inhibitors or treatments. Secretion is also blocked in a physiologically identical manner by the tyrosine phosphatase inhibitors pervanadate and bisperoxo(1,10-phenanthroline)-oxovanadate. Inhibition of either the kinase(s) or phosphatase(s) leads to the accumulation of albumin between the trans-Golgi and the plasma membrane, whereas the immediate precursor proalbumin builds up in a proximal compartment. The trans-Golgi marker TGN38 is rapidly dispersed under conditions that inhibit tyrosine phosphatase action, whereas the distribution of the cis-Golgi marker GM130 is insensitive to genistein or pervanadate. By using a specifically reactive biotinylation probe, we detected protein tyrosine phosphatases in highly purified rat liver Golgi membranes. These membranes also contain both endogenous tyrosine kinases and their substrates, indicating that enzymes and substrates for reversible tyrosine phosphorylation are normal membrane-resident components of this trafficking compartment. In the absence of perturbation of actin filaments and microtubules, we conclude that reversible protein tyrosine phosphorylation in the trans-Golgi network is essential for albumin secretion and propose that the constitutive secretion of albumin is in fact a regulated process.
Webb
Rachel J
RJ
Dept. of Physiology, University College London, London WC1E 6JJ, UK.
Judah
Jacob D
JD
Lo
Lee-Chiang
LC
Thomas
Geraint M H
GM
D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D021381:Protein Transport / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D012709:Serum Albumin / Q000378:metabolism*; D013379:Substrate Specificity; D014443:Tyrosine / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology; D021601:trans-Golgi Network / Q000187:drug effects / Q000378:metabolism*
D015415:Biomarkers; D004791:Enzyme Inhibitors; D012709:Serum Albumin; C076715:pervanadate; D014638:Vanadates; D014443:Tyrosine; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D010749:Phosphoprotein Phosphatases