Studies on methemoglobin reductase. Immunochemical similarity of soluble methemoglobin reductase and cytochrome b5 of human erythrocytes with NADH-cytochrome b5 reductase and cytochrome b5 of rat liver microsomes

Fumio Kuma, Russell A. Prough, Bettie Sue Siler Masters

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

An antibody preparation elicited against purified, lysosomal-solubilized NADH-cytochrome b5 reductase from rat liver microsomes was shown to interact with methemoglobin reductase of human erythrocytes by inhibiting the rate of erythrocyte cytochrome b5 reduction by NADH. The ferricyanide reductase activity of the enzyme was not inhibited by the antibody, suggesting that the inhibition of methemoglobin reductase activity may be due to interference with the binding of cytochrorme b5 to the flavoprotein. Under conditions of limiting concentrations of flavoprotein, the antibody inhibited the rate of methemoglobin reduction in a reconstituted system consisting of homogeneous methemoglobin reductase and cytochrome b5 from human erythrocytes. This inhibition was due to the decreased level of reduced cytochrome b5 during the steady state of methemoglobin reduction while the rate of methemoglobin reduction per reduced cytochrome b5 stayed constant, suggesting that the enzyme was not concerned with an electron transport between the reduced cytochrome b5 and methemoglobin. An antibody to purified, trypsin-solubilized cytochrome b5 from rat liver microsomes was shown to inhibit erythrocyte cytochrome b5 reduction by methemoglobin reductase and NADH to a lesser extent than microsomal cytochrome b5 preparations from rat liver (trypsin solubilized or detergent solubilized) and pig liver (trypsin solubilized). The results presented establish that soluble methemoglobin reductase and cytochrome b5 of human erythrocytes are immunochemically similar to NADH-cytochrome b5 reductase and cytochrome b5 of liver microsomes, respectively.

Original languageEnglish (US)
Pages (from-to)600-607
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume172
Issue number2
DOIs
StatePublished - 1976
Externally publishedYes

Fingerprint

Cytochrome-B(5) Reductase
Cytochromes b5
Liver Microsomes
Liver
Rats
Erythrocytes
Methemoglobin
Trypsin
Flavoproteins
Antibodies
Enzymes
Electron Transport
Detergents
NAD
Swine

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

@article{f20504d3557f441f80ccb4fd4f62db09,
title = "Studies on methemoglobin reductase. Immunochemical similarity of soluble methemoglobin reductase and cytochrome b5 of human erythrocytes with NADH-cytochrome b5 reductase and cytochrome b5 of rat liver microsomes",
abstract = "An antibody preparation elicited against purified, lysosomal-solubilized NADH-cytochrome b5 reductase from rat liver microsomes was shown to interact with methemoglobin reductase of human erythrocytes by inhibiting the rate of erythrocyte cytochrome b5 reduction by NADH. The ferricyanide reductase activity of the enzyme was not inhibited by the antibody, suggesting that the inhibition of methemoglobin reductase activity may be due to interference with the binding of cytochrorme b5 to the flavoprotein. Under conditions of limiting concentrations of flavoprotein, the antibody inhibited the rate of methemoglobin reduction in a reconstituted system consisting of homogeneous methemoglobin reductase and cytochrome b5 from human erythrocytes. This inhibition was due to the decreased level of reduced cytochrome b5 during the steady state of methemoglobin reduction while the rate of methemoglobin reduction per reduced cytochrome b5 stayed constant, suggesting that the enzyme was not concerned with an electron transport between the reduced cytochrome b5 and methemoglobin. An antibody to purified, trypsin-solubilized cytochrome b5 from rat liver microsomes was shown to inhibit erythrocyte cytochrome b5 reduction by methemoglobin reductase and NADH to a lesser extent than microsomal cytochrome b5 preparations from rat liver (trypsin solubilized or detergent solubilized) and pig liver (trypsin solubilized). The results presented establish that soluble methemoglobin reductase and cytochrome b5 of human erythrocytes are immunochemically similar to NADH-cytochrome b5 reductase and cytochrome b5 of liver microsomes, respectively.",
author = "Fumio Kuma and Prough, {Russell A.} and Masters, {Bettie Sue Siler}",
year = "1976",
doi = "10.1016/0003-9861(76)90113-2",
language = "English (US)",
volume = "172",
pages = "600--607",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Studies on methemoglobin reductase. Immunochemical similarity of soluble methemoglobin reductase and cytochrome b5 of human erythrocytes with NADH-cytochrome b5 reductase and cytochrome b5 of rat liver microsomes

AU - Kuma, Fumio

AU - Prough, Russell A.

AU - Masters, Bettie Sue Siler

PY - 1976

Y1 - 1976

N2 - An antibody preparation elicited against purified, lysosomal-solubilized NADH-cytochrome b5 reductase from rat liver microsomes was shown to interact with methemoglobin reductase of human erythrocytes by inhibiting the rate of erythrocyte cytochrome b5 reduction by NADH. The ferricyanide reductase activity of the enzyme was not inhibited by the antibody, suggesting that the inhibition of methemoglobin reductase activity may be due to interference with the binding of cytochrorme b5 to the flavoprotein. Under conditions of limiting concentrations of flavoprotein, the antibody inhibited the rate of methemoglobin reduction in a reconstituted system consisting of homogeneous methemoglobin reductase and cytochrome b5 from human erythrocytes. This inhibition was due to the decreased level of reduced cytochrome b5 during the steady state of methemoglobin reduction while the rate of methemoglobin reduction per reduced cytochrome b5 stayed constant, suggesting that the enzyme was not concerned with an electron transport between the reduced cytochrome b5 and methemoglobin. An antibody to purified, trypsin-solubilized cytochrome b5 from rat liver microsomes was shown to inhibit erythrocyte cytochrome b5 reduction by methemoglobin reductase and NADH to a lesser extent than microsomal cytochrome b5 preparations from rat liver (trypsin solubilized or detergent solubilized) and pig liver (trypsin solubilized). The results presented establish that soluble methemoglobin reductase and cytochrome b5 of human erythrocytes are immunochemically similar to NADH-cytochrome b5 reductase and cytochrome b5 of liver microsomes, respectively.

AB - An antibody preparation elicited against purified, lysosomal-solubilized NADH-cytochrome b5 reductase from rat liver microsomes was shown to interact with methemoglobin reductase of human erythrocytes by inhibiting the rate of erythrocyte cytochrome b5 reduction by NADH. The ferricyanide reductase activity of the enzyme was not inhibited by the antibody, suggesting that the inhibition of methemoglobin reductase activity may be due to interference with the binding of cytochrorme b5 to the flavoprotein. Under conditions of limiting concentrations of flavoprotein, the antibody inhibited the rate of methemoglobin reduction in a reconstituted system consisting of homogeneous methemoglobin reductase and cytochrome b5 from human erythrocytes. This inhibition was due to the decreased level of reduced cytochrome b5 during the steady state of methemoglobin reduction while the rate of methemoglobin reduction per reduced cytochrome b5 stayed constant, suggesting that the enzyme was not concerned with an electron transport between the reduced cytochrome b5 and methemoglobin. An antibody to purified, trypsin-solubilized cytochrome b5 from rat liver microsomes was shown to inhibit erythrocyte cytochrome b5 reduction by methemoglobin reductase and NADH to a lesser extent than microsomal cytochrome b5 preparations from rat liver (trypsin solubilized or detergent solubilized) and pig liver (trypsin solubilized). The results presented establish that soluble methemoglobin reductase and cytochrome b5 of human erythrocytes are immunochemically similar to NADH-cytochrome b5 reductase and cytochrome b5 of liver microsomes, respectively.

UR - http://www.scopus.com/inward/record.url?scp=0017279859&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017279859&partnerID=8YFLogxK

U2 - 10.1016/0003-9861(76)90113-2

DO - 10.1016/0003-9861(76)90113-2

M3 - Article

C2 - 1259422

AN - SCOPUS:0017279859

VL - 172

SP - 600

EP - 607

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 2

ER -