Enzyme-substrate intermediate formation at lysine 329 of human deoxyhypusine synthase.

The Journal of Biological Chemistry
Edith C WolffM H Park

Abstract

Deoxyhypusine (Nepsilon-(4-aminobutyl)lysine) is the key intermediate in the posttranslational synthesis of the unique amino acid, hypusine (Nepsilon-(4-amino-2-hydroxybutyl)lysine). Deoxyhypusine synthase catalyzes the formation of deoxyhypusine by conjugation of the butylamine moiety of spermidine to the epsilon-amino group of one specific lysine residue of the eukaryotic translation initiation factor 5A (eIF-5A) precursor protein. However, in the absence of the eIF-5A precursor, catalysis involves only the NAD-dependent cleavage of spermidine to generate 1,3-diaminopropane and a putative 4-carbon amine intermediate that gives rise to Delta1-pyrroline. We have obtained evidence for a covalent enzyme-substrate intermediate that accumulates in the absence of the eIF-5A precursor. Incubation of human recombinant enzyme with [1, 8-3H]spermidine and NAD, followed by reduction with NaBH3CN, resulted in specific radiolabeling of the enzyme. The radioactive component in the reduced enzyme intermediate was identified as deoxyhypusine and was shown to occur at a single locus. The fact that labeled deoxyhypusine was found after treatment with a reducing agent suggests an intermediate with the butylamine moiety derived from spermidine at...Continue Reading

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Citations

Mar 25, 2004·Protein Expression and Purification·Jenq-Kuen HuangLisa Wen
Dec 24, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jong-Hwan ParkMyung Hee Park
Sep 28, 2015·Biochimie·Laura Itzel Quintas-GranadosMaría Elizbeth Álvarez-Sánchez
Sep 30, 2015·Molecular Microbiology·Jürgen LassakKirsten Jung
Mar 8, 2006·The FEBS Journal·Paul M J ClementMyung H Park
Jul 18, 2014·Critical Reviews in Biochemistry and Molecular Biology·Thomas E DeverByung-Sik Shin
Sep 28, 2018·The Journal of Biological Chemistry·Myung Hee Park, Edith C Wolff
Nov 3, 2009·The Journal of Biological Chemistry·Bhavna ChawlaRentala Madhubala
Mar 29, 2000·The Journal of Biological Chemistry·E C WolffM H Park

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