EurekaMag.com logo
+ Site Statistics
References:
48,253,228
Abstracts:
22,798,842
+ Resolve Accession
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
Submit PDF Full TextSubmit PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Spectral evidence for a flavine adduct in a mono alkylated derivative of pig heart lipo amide dehydrogenase


, : Spectral evidence for a flavine adduct in a mono alkylated derivative of pig heart lipo amide dehydrogenase. Journal of Biological Chemistry 251(23): 7726-7728

A derivative of the flavoprotein pig heart lipoamide dehydrogenase has been described recently, in which 1 of the 2 cysteine residues generated on reduction of the intrachain active center disulfide bridge is selectively alkylated with iodoacetamide. This monolabeled enzyme exhibits a spectrum of oxidized bound flavin. The addition of 1 mM NAD+ to this derivative at pH 8.3 causes a decrease in absorbance of approximately 50% at 448 nm, with a concomitant increase at 380 nm. These spectral changes are complete within 3 ms and are reversible. NAD+ titrations generate isosbestic points at 408, 374 and 327 nm; allowing values for the apparent dissociation constant for NAD+ and the extent of bleaching at infinite ligand to be obtained from double reciprocal plots. Between pH 6.1 and 8.8, the apparent Kd decreases from 320-35 .mu.M, whereas the extrapolated .DELTA.epsilon.448 values remain approximately constant at 1/2 .epsilon.448. Direct measurement of NAD+ binding by gel filtration at pH 8.8 indicates that the spectral changes are associated with a stoichiometry of 1.2 mol of NAD+ bound/2 mol of FAD. The modified protein is a dimer containing 1 FAD and 1 alkylated cysteine residue/subunit; the native enzyme is also dimeric. The visible spectrum of the species absorbing at 380 nm, approximated by correction for the residual oxidized FAD, shows a single maximum at 384 nm, .epsilon.384 = 8.7 mM-1cm-1. Comparison of this spectrum with that of model compounds of known structure suggests that it may represent a reversible covalent flavin adduct induced on binding NAD+.

(PDF 0-2 workdays service)

Order PDF Full Text

Close
Close

Click here to order any other PDF Full Text


Accession: 006459676

PDF Full Text: Submit


Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:
Close
Close

Other references

Terwilliger E.F.; Langhoff E.; Haseltine W.A., 1991: A new role for the nef gene of hiv. AIDS Research & Human Retroviruses 7(2): 180

Roepke, B.; Berndt, H., 1972: Vocational rehabilitation of tumor patients. Archiv für Geschwulstforschung 39(2): 137-152

Yukich, J.O.; Taylor, C.; Eisele, T.P.; Reithinger, R.; Nauhassenay, H.; Berhane, Y.; Keating, J., 2013: Travel history and malaria infection risk in a low-transmission setting in Ethiopia: a case control study. Malaria remains the leading communicable disease in Ethiopia, with around one million clinical cases of malaria reported annually. The country currently has plans for elimination for specific geographic areas of the country. Human movement may lea...

Clarke, E.; Jenkins, N.; Nelson, R.; Ritchie, H.; Rice, G.; Singer, J.W., 1994: CT-2584, a novel anti-tumor agent, lacking apparent hematopoietic toxicity in vivo. Blood 84(10 SUPPL 1): 52A

Pellerin Louise; Alumbaugh David L., 1997: Geophysical tools for electromagnetic characterization of buried waste. Butsuri-Tansa = Geophysical Exploration 50(6): 656-663

Sidor-Smaga, M., 1968: Trial evaluation of BCG test in patients with pulmonary tuberculosis. II. Gruzlica i Choroby Pluc; Tuberculosis et Pneumonologia 36(9): 789-794

Akiyama, T.; Mizuta, T.; Kawazoe, S.; Eguchi, Y.; Kawaguchi, Y.; Takahashi, H.; Ozaki, I.; Fujimoto, K., 2011: Body mass index is associated with age-at-onset of HCV-infected hepatocellular carcinoma patients. To identify factors associated with the age at onset of hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). Five hundred and fifty-six consecutive patients positive for HCV antibody and treatment-naïve HCC diagnosed between 1995 and 2...

Ouali, S.; Ben Salem, H.; Hammas, S.; Neffeti, E.; Remedi, F.; Mahdhaoui, A.; Boughzela, E.; Mankai, R., 2011: Prevalence of Brugada-type ECG pattern and early ventricular repolarization pattern in Tunisian athletes. No data regarding the prevalence of the Brugada-type electrocardiogram (ECG) pattern and the early ventricular repolarization pattern (ERP) in the North African population were available. The aims of this study were to determine the frequency of B...

Kurosawa, Y., 1979: On the genus Thymedes Waterhouse (Coleoptera, Buprestidae) endemic to the Philippines. Bulletin National Science Museum Series A: Zoology 5(1): 51-60

Outka D.; Safa A.R., 1980: Induction of alternate life cycle stages and determination of ploidy in the coccolithophorid hymenomonas carterae. Journal of Protozoology 27(3): 13A