Molecular cloning of the mature NAD+-dependent succinic semialdehyde dehydrogenase from rat and human

cDNA isolation, evolutionary homology, and tissue expression

Ken L. Chambliss, Deborah L. Caudle, Debra D. Hinson, Carolyn R. Moomaw, Clive A. Slaughter, Cornelis Jakobs, K. Michael Gibson

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Three rat brain cDNA clones ∼3500, 1465, and 1135 base pairs in length encoding succinic semialdehyde dehydrogenase (SSADH; EC 1.2.1.24) were isolated from two cDNA libraries using a polymerase chain reaction derived probe. Restriction mapping and DNA sequencing revealed that the 3.5-kilobase clone contained an 84-base pair (28 amino acid) insert in the coding region. Composite clones encoding mature SSADH predicted proteins with 488 amino acids (Mr = 52,188) when including the insert and 460 amino acids (Mr = 48,854) without the insert. The cDNA clones were confirmed by expression of enzyme activity in bacteria and protein sequence data obtained from sequencing purified rat brain SSADH. Two human liver SSADH cDNA clones of 1091 and 899 base pairs were also isolated. Human and rat SSADH share 83 and 91% identity in nucleotide and protein sequence, respectively. Northern blot analysis revealed two differentially expressed SSADH transcripts of approximately 2.0 and 6.0 kilobases in both rat and human tissues. Human genomic Southern blots indicate that the two SSADH transcripts are encoded by a greater than 20-kilobase single copy gene. Mammalian SSADH contains significant homology to bacterial NADP+-succinic semialdehyde dehydrogenase (EC 1.2.1.16) and conserved regions of general aldehyde dehydrogenases (EC 1.2.1.3), suggesting it is a member of the aldehyde dehydrogenase superfamily of proteins.

Original languageEnglish (US)
Pages (from-to)461-467
Number of pages7
JournalJournal of Biological Chemistry
Volume270
Issue number1
DOIs
StatePublished - Jan 6 1995
Externally publishedYes

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Succinate-Semialdehyde Dehydrogenase
Cloning
Molecular Cloning
Rats
Complementary DNA
Clone Cells
Tissue
Aldehyde Dehydrogenase
Base Pairing
Amino Acids
Brain
Proteins
Die casting inserts
Polymerase chain reaction
Enzyme activity
Restriction Mapping
NADP
Gene Library
Liver
Bacteria

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Molecular cloning of the mature NAD+-dependent succinic semialdehyde dehydrogenase from rat and human : cDNA isolation, evolutionary homology, and tissue expression. / Chambliss, Ken L.; Caudle, Deborah L.; Hinson, Debra D.; Moomaw, Carolyn R.; Slaughter, Clive A.; Jakobs, Cornelis; Gibson, K. Michael.

In: Journal of Biological Chemistry, Vol. 270, No. 1, 06.01.1995, p. 461-467.

Research output: Contribution to journalArticle

Chambliss, Ken L. ; Caudle, Deborah L. ; Hinson, Debra D. ; Moomaw, Carolyn R. ; Slaughter, Clive A. ; Jakobs, Cornelis ; Gibson, K. Michael. / Molecular cloning of the mature NAD+-dependent succinic semialdehyde dehydrogenase from rat and human : cDNA isolation, evolutionary homology, and tissue expression. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 1. pp. 461-467.
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