Molecular cloning of human cDNA with a sequence highly similar to that of the dihydrofolate reductase gene in brain libraries derived from Alzheimer's disease patients

Eur J Biochem. 1995 May 15;230(1):337-43. doi: 10.1111/j.1432-1033.1995.0337i.x.

Abstract

A polyclonal antibody was raised against a serine protease purified from the extracellular fluid of familial Alzheimer's disease lymphoblastoid cells. Using this antibody, a cDNA library from familial Alzheimer's disease cells and two cDNA libraries from the brains of two Alzheimer's disease patients were screened independently. The familial Alzheimer's disease protein 1 (FADP1) cDNA clones isolated from these three libraries were subjected to DNA sequence analysis. The nucleotide sequence of FADP1 cDNA is highly similar to the 5' portion of the human dihydrofolate reductase (DHFR) gene, however, the sequence corresponding to exon 1 of the DHFR gene is completely disrupted and contains a 247-bp DNA insert with a sequence unique to FADP1. Moreover, FADP1 cDNA harbours a large open reading frame, including the unique insert, which has the potential to code an approximately 50-kDa protein. The deduced amino acid sequence of this protein contains 12 cysteine residues potentially involved in six disulfide bonds, a proline-rich segment and a hydrophobic segment. Northern-blot analysis with the unique insert DNA probe verified that FADP1 protein is expressed in both lymphoblastoid and brain cells derived from Alzheimer's disease patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Aged
  • Alzheimer Disease / metabolism*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Brain / enzymology*
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • Female
  • Gene Library
  • Humans
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Proteins / chemistry
  • Proteins / genetics*
  • RNA, Messenger / analysis
  • Rabbits
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / genetics*
  • Tetrahydrofolate Dehydrogenase / genetics*

Substances

  • DNA, Complementary
  • Proteins
  • RNA, Messenger
  • familial Alzheimer's disease protein 1
  • Tetrahydrofolate Dehydrogenase
  • Serine Endopeptidases

Associated data

  • GENBANK/X81004