Incorporation and distribution of selenium into thiolase from Clostridium kluyveri.
Clostridium kluyveri incorporates selenium as selenomethionine into its acetoacetyl-CoA thiolase when grown in media containing normal sulfur-to-selenium ratios. Antibodies raised against the purified enzyme permitted quantitative immunoprecipitation of thiolase from crude cell extracts and thus facilitated the systematic analysis of the effects of wide variation in sulfur-to-selenium ratios on selenium incorporation into the enzyme. The extent of incorporation of selenium into thiolase was found to be dependent on the form of selenium supplied. When [75Se]selenomethionine was the source of selenium, the incorporation of selenium into thiolase was inversely proportional to the level of added methionine. However, similar levels of methionine failed to decrease the incorporation of selenium from selenite. To study the location of selenomethionine and methionine residues in the polypeptide chain of the enzyme, thiolase was prepared from cells cultured in the presence of H2 35SO4 or Na2 75SeO3. The 35S- or 75Se-labeled protein was treated with trypsin and the resulting peptides were isolated by reverse phase high performance liquid chromatography. The peptide maps of the enzyme indicated that selenium was distributed throughout the primary structure in a manner that paralleled methionine. From these studies, it is concluded that selenium occurs in thiolase adventitiously and is not required for any biological function.
Sliwkowski
M X
MX
Stadtman
T C
TC
D000101:Acetyl-CoA C-Acetyltransferase / Q000378:metabolism*; D000123:Acetyltransferases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007163:Immunosorbent Techniques; D008715:Methionine / Q000378:metabolism; D012643:Selenium / Q000378:metabolism*; D013455:Sulfur / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D013455:Sulfur; D008715:Methionine; D000123:Acetyltransferases; D000101:Acetyl-CoA C-Acetyltransferase; D014357:Trypsin; D012643:Selenium