Abstract

Protein disulfide isomerase (PDI) catalyzed the interchange of disulfide bonds in substrate proteins. The two Cys-Gly-His-Cys active sites provide a thiol of low pKa and a disulfide bond of high reduction potential (E??). Trans-1,20bis(2-mercaptoacetamido)cyclohexane (BMC) mimics the pKa and E?? of the PDI active sites. In vitro assays demonstrate that BMC is an efficient catalyst of the reactivation of scrambled ribonuclease (Rnase) A, a substrate with non-native disulfide bonds. In vivo assays show that BMC in the growth medium of yeast cells increases the secretion of native acid phosphatase, which has eight disulfide bonds. The effect is similar to that from the overproduction of PDI in the yeast cells. A monothiol analog of BMC, N-methyl-mercaptoacetamide (NMA), also catalyzes the reactivation of scrambled Rnase A, but less efficiently than does BMC. These data indicate that molecules with a low thiol pKa and high disulfide E?? catalyze effectively the formation of native disulfide bonds in proteins. Moreover, BMC is a useful catalyst for the in vitro and in vivo folding of proteins that contain multiple disulfide bonds.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002301-18
Application #
6575312
Study Section
Project Start
2002-03-01
Project End
2003-02-28
Budget Start
Budget End
Support Year
18
Fiscal Year
2002
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

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Wijayatunga, Nadeeja N; Sams, Valerie G; Dawson, John A et al. (2018) Roux-en-Y gastric bypass surgery alters serum metabolites and fatty acids in patients with morbid obesity. Diabetes Metab Res Rev 34:e3045
Assadi-Porter, Fariba M; Reiland, Hannah; Sabatini, Martina et al. (2018) Metabolic Reprogramming by 3-Iodothyronamine (T1AM): A New Perspective to Reverse Obesity through Co-Regulation of Sirtuin 4 and 6 Expression. Int J Mol Sci 19:
Dominguez, Eddie; Zarnowski, Robert; Sanchez, Hiram et al. (2018) Conservation and Divergence in the Candida Species Biofilm Matrix Mannan-Glucan Complex Structure, Function, and Genetic Control. MBio 9:
Franco, Aldo; Dovell, Sanaz; Möller, Carolina et al. (2018) Structural plasticity of mini-M conotoxins - expression of all mini-M subtypes by Conus regius. FEBS J 285:887-902
Wales, Jessica A; Chen, Cheng-Yu; Breci, Linda et al. (2018) Discovery of stimulator binding to a conserved pocket in the heme domain of soluble guanylyl cyclase. J Biol Chem 293:1850-1864
Selen Alpergin, Ebru S; Bolandnazar, Zeinab; Sabatini, Martina et al. (2017) Metabolic profiling reveals reprogramming of lipid metabolic pathways in treatment of polycystic ovary syndrome with 3-iodothyronamine. Physiol Rep 5:
Mong, Surin K; Cochran, Frank V; Yu, Hongtao et al. (2017) Heterochiral Knottin Protein: Folding and Solution Structure. Biochemistry 56:5720-5725

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