Abstract
Highly purified molybdoferredoxin, with a specific activity of 2.6 mumoles of acetylene reduced per min per mg of protein, was obtained from Clostridium pasteurianum. The protein at concentrations above 5 mg/ml exists in solution as a tetrameric complex with two subunits each of about 60,000 and 50,000 daltons. Two atoms of molybdenum are present per protein molecule of 220,000 daltons. The S(0) (20, w) was found to be 10.5. The tetramer dissociates into a dimer as demonstrated by a decreasing sedimentation coefficient with decreasing protein concentration. At low pH and ionic strength, further dissociation into the monomers is achieved. A method for the isolation of the protein subunits is described.
MeSH Terms
Acetylene/metabolism
Bacterial Proteins/analysis,isolation & purification,metabolism
Chromatography, DEAE-Cellulose
Clostridium/analysis
Colorimetry
Densitometry
Electrophoresis, Polyacrylamide Gel
Ferredoxins/analysis,isolation & purification,metabolism
Hydrogen-Ion Concentration
Molecular Weight
Molybdenum/analysis,isolation & purification,metabolism
Organometallic Compounds/analysis,isolation & purification,metabolism
Protein Conformation
Sodium Dodecyl Sulfate
Ultracentrifugation
Chemicals
Bacterial Proteins
Ferredoxins
Organometallic Compounds
Sodium Dodecyl Sulfate
Molybdenum
Acetylene
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Huang T C
Zumft W G
Mortenson L E
References (13)
13 references, click to expand
-
The gel-filtration behaviour of proteins related to their molecular weights over a wide range.
Biochem J. 1965 Sep;96(3):595-606
PMID: 5862401
-
Properties of azoferredoxin purified from nitrogen-fixing extracts of Clostridium pasteurianum.
Biochim Biophys Acta. 1969 Jan 14;172(1):106-15
PMID: 5763411
-
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.
J Biol Chem. 1969 Aug 25;244(16):4406-12
PMID: 5806584
-
The estimation of polypeptide chain molecular weights by gel filtration in 6 M guanidine hydrochloride.
J Biol Chem. 1969 Sep 25;244(18):4989-94
PMID: 5824574
-
Purification and properties of the constituents of the nitrogenase complex from Clostridium pasteurianum.
J Bacteriol. 1970 Mar;101(3):794-801
PMID: 5438048
-
The oxygen sensitivity of spinach ferredoxin and other iron-sulfur proteins. The formation of protein-bound sulfur-zero.
J Biol Chem. 1971 Feb 10;246(3):643-53
PMID: 5542679
-
Molecular weight and subunit structure of molybdoferredoxin from Clostridium pasteurianum W5.
Biochim Biophys Acta. 1971 Feb 16;229(2):431-6
PMID: 5553986
-
Purification and some properties of molybdoferredoxin, a component of nitrogenase from Clostridium pasteurianum.
Biochemistry. 1971 May 25;10(11):2066-72
PMID: 4327398
-
On the structure and function of nitrogenase from Clostridium pasteurianum W5.
Biochem Biophys Res Commun. 1972 Sep 26;48(6):1525-32
PMID: 4342714
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
A micro biuret method for protein determination; determination of total protein in cerebrospinal fluid.
Scand J Clin Lab Invest. 1953;5(3):218-22
PMID: 13135413
-
STUDIES ON THE CHEMICAL NATURE OF CLOSTRIDIAL FERREDOXIN.
J Biol Chem. 1963 Dec;238:3899-913
PMID: 14086723
-
THE CONTENT AND POSSIBLE CATALYTIC SIGNIFICANCE OF LABILE SULFIDE IN SOME METALLOFLAVOPROTEINS.
J Biol Chem. 1965 May;240:2222-8
PMID: 14299651