+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Probing the active site of the reconstituted carnitine carrier from rat liver mitochondria with sulfhydryl reagents. A cysteine residue is localized in or near the substrate binding site

, : Probing the active site of the reconstituted carnitine carrier from rat liver mitochondria with sulfhydryl reagents. A cysteine residue is localized in or near the substrate binding site. European Journal of Biochemistry 228(2): 271-278

The interaction of sulfhydryl reagents with the carnitine carrier of rat liver mitochondria was studied in detail in proteoliposomes. The addition of N-ethylmaleimide, mercurials at low concentrations, Cu(2+)-phenanthroline and diamide modified a single sulfhydryl group (the class II group) that is involved in transport function. The treatment of the inhibited protein with 1,4-dithioerythritol led to full recovery of carnitine exchange except for N-ethylmaleimide. Evidence is provided that the addition of carnitine to the carrier blocks the interaction of the sulfhydryl reagents with the protein. This result strongly suggests that the critical cysteine residue is localized in, or near, the substrate binding site. Interaction of other cysteine residues in the carrier protein with high concentrations of mercurials modified another class of sulfhydryl groups (the class I group) that are not directly involved in carnitine transport. The oxidized and reduced forms of the carnitine carrier show slightly different molecular masses on SDS/PAGE. Disulfide bridge(s) induced by Cu(2+)-phenanthroline and diamide are present in a single polypeptide part of the protein and induced no disulfide bridges between two polypeptide chains.


PMID: 7705339

DOI: 10.1111/j.1432-1033.1995.0271n.x

Other references

Sivadjian, J., 1951: Hygro-photographic study of transpiration in plants. Comptes Rendus Hebdomadaires des Seances de L'academie des Sciences 232(21): 1956-1958

Rabinkova E.V.; Torosyan M.V.; Shishkova O.V., 1983: Effect of r 6k plasmid on cell resistance to radiation in escherichia coli k 12. The effect of R6K plasmid on the UV- and .gamma.-ray sensitivity of E. coli K-12 strains was studied. The plasmid-mediated protection was shown to be expressed only in recA, lexA, recB, uvrA mutants, but not in the wild type AB 1157 strain. The pr...

Sammour M.B.; Youssef H.H.; Said A.; Abd E.S.lam A.F.; E.H.ig M.O., 1977: Ovarian activity in bilharzial hepatic fibrosis. Ain Shams Medical Journal 28(1/2): 25-30

Kuznetsova, S.M., 1982: Erythromycin in the modern antibiotic therapy of bacterial infections (a review lecture). Antibiotiki 27(5): 365-372

Clayton, D.A., 1987: Nuclear gene products that function in mitochondrial DNA replication. Mammalian mitochondrial DNAs replicate unidirectionally from two distinct strand-specific origins. A round of replication begins at the heavy-strand origin (the D-loop) where transcripts from an upstream promoter serve as the primers for DNA synth...

Altea, E., 1950: Effect of the human cerebrospinal fluid on the activity of the cholinesterase. Bollettino Della Societa Italiana di Biologia Sperimentale 26(3): 236-238

Gyorgy, E.B., 1984: Occurrence of Diaporthe helianthi (Phomopsis helianthi Munt. Cvet. et al.) in the Tolna district. The fungus was isolated from sunflower plants with symptoms of a new stalk disease.

Liu, Y.; Xu, G.; Wei, J.; Wu, Q.; Li, X., 2017: Cardiomyocyte coculture on layered fibrous scaffolds assembled from micropatterned electrospun mats. Challenges remain in engineering cardiac tissues with functional and morphological properties similar to those of native myocardium. In the current study, micropatterned fibrous mats are obtained by deposition of electrospun fibers on lithographic...

Smolen, J.M.; Stone, A.T., 1998: Metal (hydr)oxide surface-catalyzed hydrolysis of chlorpyrifos-methyl, chlorpyrifos-methyl oxon, and paraoxon. This work is concerned with the susceptibility of pesticides and their transformation products to mineral-surface-catalyzed hydrolysis in soils. Experiments were performed in an aqueous medium containing pH buffer [2.0-5.0 mM acetate or 3-(N-morph...

Glick S.D.; Badalamenti J.I., 1986: Sex difference in reward asymmetry and effects of cocaine. Male and female rats with bilaterally implanted electrodes in the lateral hypothalamus were tested daily for self-stimulation of each side of the brain, and rotation (circling behavior) was recorded concomitantly. All rats rotated in a preferred d...