Degradation of human thyroperoxidase in the endoplasmic reticulum involves two different pathways depending on the folding state of the protein.
Human thyroperoxidase (hTPO), a type I transmembrane glycoprotein, plays a key role in thyroid hormone synthesis. In a previous paper (Fayadat, L., Niccoli, P., Lanet, J., and Franc, J. L. (1998) Endocrinology 139, 4277-4285) we established that after the synthesis, only 15-20% of the hTPO molecules were recognized by a monoclonal antibody (mAb15) directed against a conformational structure and that only 2% were able to reach the cell surface. In the present study using pulse-chase experiments in the presence or absence of protease inhibitors followed by immunoprecipitation procedures with monoclonal antibodies recognizing unfolded or partially folded hTPO forms we show that: (i) unfolded hTPO forms are degraded by the proteasome and (ii) partially folded hTPO forms are degraded by other proteases. It was also established upon incubating endoplasmic reticulum (ER) membranes in vitro that the degradation of the partially folded hTPO was carried out by serine and cysteine integral ER membrane proteases. These data provide valuable insights into the quality control mechanisms whereby the cells get rid of misfolded or unfolded proteins. Moreover, this is the first study describing a protein degradation process involving two distinct degradation pathways (proteasome and ER cysteine/serine proteases) at the ER level, depending on the folding state of the protein.
Fayadat
L
L
U38 INSERM, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille cedex 5, France.
Siffroi-Fernandez
S
S
Lanet
J
J
Franc
J L
JL
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016466:CHO Cells; D002738:Chloroquine / Q000494:pharmacology; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004640:Emetine / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D009097:Multienzyme Complexes / Q000378:metabolism; D009195:Peroxidase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D017510:Protein Folding; D013961:Thyroid Gland / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D015853:Cysteine Proteinase Inhibitors; D009097:Multienzyme Complexes; D000643:Ammonium Chloride; C067713:lactacystin; D002738:Chloroquine; D003513:Cycloheximide; D009195:Peroxidase; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine; D004640:Emetine