Comparison of the intracellular trafficking of two alternatively spliced isoforms of pp120, a substrate of the insulin receptor tyrosine kinase

Curtis Choice, Matthew N. Poy, Pietro Formisano, Sonia M. Najjar

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

pp120, a substrate of the insulin receptor tyrosine kinase, is a plasma membrane glycoprotein in the hepatocyte. It is expressed as two spliced isoforms differing by the presence (full length) or absence (truncated) of most of the intracellular domain including all phosphorylation sites. Because the two isoforms differ by their ability to regulate receptor-mediated insulin-endocytosis and degradation, we aimed to investigate the cellular basis four this functional difference by comparing their intracellular trafficking. During its intracellular assembly, pp120 is transported from the trans-Golgi network to the-sinusoidal domain of the plasma membrane before its final transcytosis to the bile canalicular domain. Because both-isoforms are expressed in hepatocytes, we examined their intracellular trafficking in NIH 3T3 fibroblasts individually transfected with each isoform. Pulse-chase experiments demonstrated that most of the newly synthesized full-length isoform reached complete maturation at about 60 min of chase. By contrast, only about 40% of the newly synthesized truncated isoform underwent complete maturation, even at more prolonged chase. Moreover, a significant portion of the truncated isoform appeared to be targeted to lysosomes. Abolishing basal phosphorylation on Ser503 by cAMP-dependent serine kinase by mutating this residue to alanine was correlated with incomplete maturation of full length pp120 in NIH 3T3 cells and hepatocytes. This finding suggests that the intracellular domain of pp120 contains information that regulates its vectorial sorting from the trans-Golgi network to the plasma membrane.

Original languageEnglish (US)
Pages (from-to)133-142
Number of pages10
JournalJournal of Cellular Biochemistry
Volume76
Issue number1
DOIs
StatePublished - 1999
Externally publishedYes

Fingerprint

Protein Isoforms
Substrates
Cell membranes
trans-Golgi Network
Hepatocytes
Phosphorylation
Cell Membrane
Transcytosis
NIH 3T3 Cells
Protein-Serine-Threonine Kinases
Insulin Receptor
Membrane Glycoproteins
Fibroblasts
Endocytosis
Lysosomes
insulin receptor tyrosine kinase
Sorting
Bile
Alanine
Insulin

Keywords

  • Assembly
  • Intracellular synthesis
  • Kinetics
  • Pulse-chase
  • Receptor-mediated endocytosis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Comparison of the intracellular trafficking of two alternatively spliced isoforms of pp120, a substrate of the insulin receptor tyrosine kinase. / Choice, Curtis; Poy, Matthew N.; Formisano, Pietro; Najjar, Sonia M.

In: Journal of Cellular Biochemistry, Vol. 76, No. 1, 1999, p. 133-142.

Research output: Contribution to journalArticle

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