Alignment and partial structural analysis of the cyanogen bromide fragments from yeast inorganic pyrophosphatase

Journal Article · Tue Jan 01 04:00:00 EST 1974 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00698a015· OSTI ID:5263375

Sterner, R; Noyes, C; Heinrikson, R L

Yeast inorganic pyrophosphatase (EC 3.6.1.1, pyrophosphate phosphohydrolase) was cleaved by cyanogen bromide in 70% formic acid for 48 hr at room temperature, and the reaction mixture was subsequently reduced and S-carboxylmethylated. Gel filtration of the cleavage products on a column of Sephadex G-50 developed in 10% formic acid gave three major components, the compositions of which accounted for the total amino acid content of the protein monomer. The alignment of these fragments generated by cleavage at the two methionine residues in the pyrophosphatase subunit was based upon partial sequence analysis at the amino and carboxyl termini of the peptides and the intact enzyme. In addition to these fragments, three minor components were isolated in a highly purified form by the gel filtration step. These were shown to be derived from the internal CNBr fragment, possibly as a result of cleavages during prolonged exposure to 70% formic acid. Subsequent analysis of the six cleavage products by automated Edman degradation and digestion with carboxypeptidase A allowed placement of 108 of the 270 residues in the subunit of yeast inorganic pyrophosphatase.

• Research Organization: Univ. of Chicago
• OSTI ID: 5263375
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 13:1; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
BIOMASS
BIOSYNTHESIS
BROMIDES
BROMINE COMPOUNDS
CARBOXYLIC ACIDS
CYANOGEN
ENERGY SOURCES
ENZYMES
ESTERASES
FUNGI
HALIDES
HALOGEN COMPOUNDS
HYDROLASES
KINETICS
MICROORGANISMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHATASES
PHOSPHORUS COMPOUNDS
PLANTS
PYROPHOSPHATES
REACTION KINETICS
RENEWABLE ENERGY SOURCES
STRUCTURAL CHEMICAL ANALYSIS
SYNTHESIS
YEASTS