Delta3,5-delta2,4-dienoyl-CoA isomerase from rat liver. Molecular characterization

S A Filppula, A I Yagi, S H Kilpeläinen, D Novikov, D R FitzPatrick, M Vihinen, D Valle, J K Hiltunen

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

rECH1, a recently identified rat cDNA (FitzPatrick, D. R., Germain-Lee, E., and Valle, D. (1995) Genomics 27, 457-466) encodes a polypeptide belonging to the hydratase/isomerase superfamily. We modeled the structure of rECH1 based on rat mitochondrial 2-enoyl-CoA hydratase 1. The model predicts that rECH1p has the hydratase fold in the core domain and two domains for interaction with other subunits. When we incubated 3,5,8,11, 14-eicosapentaenoyl-CoA with purified rECH1p, the spectral data suggested a switching of the double bonds from the Delta3-Delta5 to the Delta2-Delta4 positions. This was confirmed by demonstrating that the product was a valid substrate for 2,4-dienoyl-CoA reductase. These results indicate that rECH1p is Delta3,5-Delta2,4-dienoyl-CoA isomerase. Subcellular fractionation and immunoelectron microscopy using antibodies to a synthetic polypeptide derived from the C terminus of rECH1p showed that rECH1p is located in the matrix of both mitochondria and peroxisomes in rat liver. Consistent with these observations, the 36,000-Da rECH1p has a potential N-terminal mitochondrial targeting signal as well as a C-terminal peroxisomal targeting signal type 1. Transport of the protein into the mitochondria with cleavage of the targeting signal results in a mature mitochondrial form with a molecular mass of 32,000 Da; transport to peroxisomes yields a protein of 36,000 Da.
Original languageEnglish
Pages (from-to)349-55
Number of pages7
JournalJournal of Biological Chemistry
Volume273
Issue number1
Publication statusPublished - 2 Jan 1998

Keywords / Materials (for Non-textual outputs)

  • Amino Acid Sequence
  • Animals
  • Carbon-Carbon Double Bond Isomerases
  • Kinetics
  • Microscopy, Immunoelectron
  • Mitochondria, Liver
  • Models, Molecular
  • Molecular Sequence Data
  • Rats
  • Sequence Homology, Amino Acid
  • Subcellular Fractions

Fingerprint

Dive into the research topics of 'Delta3,5-delta2,4-dienoyl-CoA isomerase from rat liver. Molecular characterization'. Together they form a unique fingerprint.

Cite this