PMID: 9543006May 23, 1998Paper

Effects of sulfhydryl regents on the activity of lambda Ser/Thr phosphoprotein phosphatase and inhibition of the enzyme by zinc ion

Protein Engineering
S Zhuo, J E Dixon

Abstract

Sulfhydryl reagents, such as dithiothreitol (DTT), affected the activity of Ser/Thr phosphoprotein phosphatases. Addition of DTT to the assay buffer increased the affinity of lambda Ser/Thr phosphoprotein phosphatase (lambda-PPase) for its Mn2+ cofactor. On the other hand, the enzyme was found to be inactivated simply by dilution in Tris buffer. The inactivation could be completely prevented by the presence of DTT or Mn2+ in the buffer. Further studies showed that oxidation or reduction of cysteine residues in lambda-PPase may not be the cause of the change in the enzyme activity. Without exception, mutation of all cysteine residues in lambda-PPase to serine did not convert the enzyme into a thiol-insensitive mutant. By careful examination of the effects of different sulfhydryl reagents, metal ion cofactors and substrates on lambda-PPase, it was found that the role of sulfhydryl reagents was the chelation of small amounts of inhibitory metal ions, which were present in plastic laboratory ware, such as disposable cuvets and tubes, with prevention of the enzyme from inactivation. One of the main contaminants found in plastic cuvets was Zn2+, which is a potent inhibitor of lambda-PPase. The inhibition of lambda-PPase by Zn2+ was c...Continue Reading

Citations

Nov 22, 2001·Archives of Biochemistry and Biophysics·A V AndreevaM A Kutuzov
Mar 8, 2011·American Journal of Physiology. Lung Cellular and Molecular Physiology·Paula J BernalClaudette M St Croix
Apr 27, 2001·Biochemical and Biophysical Research Communications·M A Kutuzov, A V Andreeva
Feb 22, 2008·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Akihiro NakayamaHiromu Sakurai
Oct 26, 2010·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Savvas N GeorgiadesRamon Vilar
Oct 19, 2010·Toxicology and Applied Pharmacology·Hajo HaaseLothar Rink
Jul 15, 2015·The Journal of Nutritional Biochemistry·Johnathan R NuttallPatricia I Oteiza
Mar 5, 2010·European Journal of Immunology·Jennifer KaltenbergHajo Haase
Apr 10, 2015·Neuroscience Bulletin·Yan XiongRong Liu
Nov 30, 2019·Proceedings of the National Academy of Sciences of the United States of America·Sun A KimMary Lou Guerinot
Mar 5, 2021·Redox Biology·Christopher Hübner, Hajo Haase
Dec 26, 2001·Chemical Reviews·A Y Louie, T J Meade
Apr 7, 2006·Analytical Biochemistry·Peter StenlundOlof P Karlsson

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