Binding of acrylonitrile to parvalbumin.
A previous study has shown that acrylonitrile (ACN) has a long half-life in rainbow trout muscle and that [14C]ACN appears to be bound to a 10,000-Da protein in muscle. The labeled protein was purified from muscle of trout exposed to [14C]ACN, separated on 20% SDS-PAGE, and digested for amino acid analysis and sequence analysis. These studies indicated that the labeled protein was the Ca(2+)-binding protein parvalbumin. Parvalbumin is an important calcium-binding protein thought to be involved in the regulation of calcium levels in various parts of the body ranging from neurons to fast-twitch muscle contractions. To study the reaction between parvalbumin and [14C]ACN, frog parvalbumin was incubated with [14C]ACN in vitro under various conditions. These studies indicated that the maximum labeling occurred at 1 nmol/nmol parvalbumin and at pH 7. Amino acid analysis of the labeled protein indicated that the labeled amino acid was probably histidine, and endoproteinase Glu-C (V-8) digestion studies revealed that the 14C was in the 1-81 amino acid segment of the protein, an area that contains two histidines.
Lech
J J
JJ
Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee 53226, USA.
Lewis
S K
SK
Friedman
M A
MA
Johnson
L A
LA
Mende-Mueller
L M
LM
D000181:Acrylonitrile / Q000378:metabolism* / Q000633:toxicity; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002021:Buffers; D002118:Calcium / Q000378:metabolism; D002250:Carbon Radioisotopes; D004591:Electrophoresis, Polyacrylamide Gel; D005343:Fibrinolytic Agents / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007553:Isotope Labeling; D008666:Metalloendopeptidases / Q000378:metabolism; D008969:Molecular Sequence Data; D009132:Muscles / Q000187:drug effects / Q000378:metabolism*; D017686:Oncorhynchus mykiss; D010320:Parvalbumins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011892:Rana catesbeiana; D017386:Sequence Homology, Amino Acid
D000596:Amino Acids; D002021:Buffers; D002250:Carbon Radioisotopes; D005343:Fibrinolytic Agents; D010320:Parvalbumins; D006639:Histidine; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D000181:Acrylonitrile; D002118:Calcium