Regulation of the expression of pp160, a putative insulin receptor signal protein, by insulin, dexamethasone, and 1-methyl-3-isobutylxanthine in 3T3-L1 adipocytes.
pp160, a cytosolic protein with Mr of approximately 160,000, is phosphorylated on tyrosine in response to insulin and is considered to be involved in signaling from the insulin receptor. The expression of pp160 during the differentiation of 3T3-L1 fibroblasts to adipocytes and in adipocytes has been investigated using quantitative immunoblotting with antibodies against a peptide from pp160. Between day 6 and day 8 of differentiation induced by insulin, dexamethasone (Dex), and 1-methyl-3-isobutylxanthine (Mix), pp160 expression increased 10-20-fold over the amount present in confluent fibroblasts. Omission of either insulin or Dex resulted in reduced expression of pp160 and in incomplete adipogenesis. Chronic treatment of fully differentiated adipocytes for 24 h with either insulin, Dex, or Mix alone in the presence of serum resulted in a decrease in the expression of pp160 by 70-85%. Chronic exposure to insulin caused a significant increase in the apparent size of pp160 to 172 kDa. Alkaline phosphatase treatment lowered the Mr of pp160 from both insulin-treated and basal cells to 150,000. These results demonstrate that pp160 is expressed in 3T3-L1 adipocytes during the time when insulin receptors are expressed in large numbers and that the maintenance of pp160 concentrations in adipocytes can be regulated by insulin, Mix, and Dex. The decreased expression of pp160 caused by these factors may be related to postreceptor insulin resistance.
Rice
K M
KM
Department of Biochemistry and Molecular Biology, Texas Tech University Health Sciences Center, Lubbock 79430.
Lienhard
G E
GE
Garner
C W
CW
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology*; D016475:3T3 Cells; D000273:Adipose Tissue / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D021382:Protein Sorting Signals / Q000302:isolation & purification / Q000378:metabolism*; D011972:Receptor, Insulin / Q000302:isolation & purification / Q000378:metabolism*
D007328:Insulin; D021382:Protein Sorting Signals; D003907:Dexamethasone; D011972:Receptor, Insulin; D015056:1-Methyl-3-isobutylxanthine