PMID: 11904218Mar 21, 2002Paper

Recombinant lysine:N(6)-hydroxylase: effect of cysteine-->alanine replacements on structural integrity and catalytic competence

Biochimica Et Biophysica Acta
S DickT Viswanatha

Abstract

Recombinant lysine:N(6)-hydroxylase, rIucD, catalyzes the hydroxylation of L-lysine to its N(6)-hydroxy derivative, with NADPH and FAD serving as cofactors in the reaction. The five cysteine residues present in rIucD can be replaced, individually or in combination, with alanine without effecting a major change in the thermal stability, the affinity for L-lysine and FAD, as well as the k(cat) for mono-oxygenase activity of the protein. However, when the susceptibility to modification by either 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) or 2,6-dichlorophenol indophenol (DPIP) serves as the criterion for monitoring conformational change(s) in rIucD and its muteins, Cys146-->Ala and Cys166-->Ala substitutions are found to induce an enhancement in the reactivity of one of the protein's remaining cysteine residues with concomitant diminution of mono-oxygenase function. In addition, the systematic study of cysteine-->alanine replacement has led to the identification of rIucD's Cys166 as the exposed residue which is detectable during the reaction of the protein with DTNB but not with iodoacetate. Substitution of Cys51 of rIucD with alanine results in an increase in mono-oxygenase activity (approx. 2-fold). Such replacement, unlike thos...Continue Reading

References

Apr 1, 1979·Analytical Biochemistry·P W RiddlesB Zerner
Jan 1, 1979·Advances in Protein Chemistry·P L Privalov
Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Jul 11, 1979·Biochimica Et Biophysica Acta·M A ParniakT Viswanatha
Feb 1, 1986·Journal of Bacteriology·V de LorenzoJ B Neilands
Dec 1, 1971·Analytical Biochemistry·G TomlinsonT Viswanatha
Jan 1, 1984·Molecular & General Genetics : MGG·R GrossV Braun
Jan 1, 1995·Sub-cellular Biochemistry·J M Sanchez-Ruiz
Nov 10, 1993·Biochimica Et Biophysica Acta·A M ThariathT Viswanatha
Jul 1, 1996·Archives of Biochemistry and Biophysics·A J TrimboliM J Barber
Jun 6, 2000·Journal of Protein Chemistry·S DickT Viswanatha
Oct 1, 1958·Biochimica Et Biophysica Acta·V MASSEY
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN
Sep 1, 1963·Biochemistry·H I HADLER, M J ERWIN

❮ Previous
Next ❯

Citations

Apr 9, 2009·Biochemical and Biophysical Research Communications·Dave Gardonio, Stefan Siemann
Dec 11, 2003·The Journal of Biological Chemistry·Alejandra YepJack Preiss
May 23, 2002·The Journal of Biological Chemistry·P J BrittoJ Wolff

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.