Protein disulfide isomerase appears necessary to maintain the catalytically active structure of the microsomal triglyceride transfer protein.
Protein disulfide isomerase (PDI) is a component of the microsomal triglyceride transfer protein (MTP) complex. This study was initiated to help elucidate the role of PDI in MTP. The 88-kDa polypeptide of MTP (88K) was dissociated from PDI by using chaotropic agents (NaClO4 and KSCN), low concentrations of a denaturant (guanidine hydrochloride) or a nondenaturing detergent (octyl glucoside). As assessed by fluorescence and circular dichroism spectroscopy, these three different approaches appeared to dissociate the components of MTP under mild, nondenaturing conditions. The dissociating agents were diluted or removed by dialysis, and the free PDI and 88K were further characterized. In all cases, the dissociation coincided with the loss of triglyceride transfer activity. The free 88-kDa polypeptide readily aggregated, suggesting that it is a hydrophobic peptide. Even in the presence of chaotropic agents, when 88K was not aggregated, transfer activity was not expressed. These results suggest that the association of PDI with 88K is necessary to maintain the catalytically active form of the triglyceride transfer protein and prevent the aggregation of 88K.
Wetterau
J R
JR
Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Ohio 45267-0575.
Combs
K A
KA
McLean
L R
LR
Spinner
S N
SN
Aggerbeck
L P
LP
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000187:drug effects; D002384:Catalysis; D002417:Cattle; D053480:Cholesterol Ester Transfer Proteins; D003902:Detergents; D006023:Glycoproteins; D006146:Guanidines / Q000494:pharmacology; D007535:Isomerases / Q000737:chemistry / Q000494:pharmacology*; D008862:Microsomes, Liver / Q000737:chemistry* / Q000187:drug effects / Q000201:enzymology; D008970:Molecular Weight; D011489:Protein Denaturation / Q000187:drug effects; D019704:Protein Disulfide-Isomerases; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D014280:Triglycerides / Q000737:chemistry*
D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D003902:Detergents; D006023:Glycoproteins; D006146:Guanidines; D014280:Triglycerides; D007535:Isomerases; D019704:Protein Disulfide-Isomerases