PMID- 10491199
OWN - NLM
STAT- MEDLINE
DCOM- 19991122
LR  - 20190620
IS  - 0014-2956 (Print)
IS  - 0014-2956 (Linking)
VI  - 265
IP  - 1
DP  - 1999 Oct 1
TI  - Human TRH-degrading ectoenzyme cDNA cloning, functional expression, genomic
      structure and chromosomal assignment.
PG  - 415-22
AB  - Thyrotropin-Releasing Hormone (TRH) is an important extracellular signal
      substance that acts as a stimulator of hormone secretion from adenohypophyseal
      target cells and fulfills many criteria for the function of a
      neuromodulator/neurotransmitter within the central and peripheral nervous
      systems. The inactivation of TRH-signals is catalysed by a highly specific
      ectoenzyme. Here, we characterize the human TRH-degrading ectoenzyme (TRH-DE) by 
      primary sequence, functional expression, genomic structure and chromosomal
      assignment. By screening a cDNA-library constructed from human lung, 5.7 kb of
      cDNA were identified. The longest open reading frame predicts a type II integral 
      membrane protein of 117 kDa. The extracellular domain contains the HEXXH + E
      motif that is characteristic of a certain family of Zn-dependent aminopeptidases.
      Within this family, the sequences of human and rat TRH-DE reveal an unusual high 
      degree of conservation (96% identical residues). Specific enzymatic activity was 
      observed after transfecting COS-7 cells with human TRH-DE cDNA yielding a Km for 
      TRH hydrolysis of 29.7 microM. Northern blot analysis demonstrated a restricted
      tissue distribution with highest transcript levels in the brain. Using
      fluorescent in situ hybridization with the cDNA and a genomic lambda clone,
      respectively, we localized the TRH-DE gene to the long arm of human chromosome
      12. Five independent P1 artificial chromosome clones were required to span the
      complete cDNA sequence and revealed that it is distributed on 19 exons.
      Interspecies Southern analysis suggests that the gene is present as a single copy
      in human, monkey, rat, mouse, dog, bovine, rabbit and chicken DNA. All of these
      data further the notion that the TRH-DE is not an ordinary enzyme but a specific 
      neuropeptidase that has been highly conserved among species.
FAU - Schomburg, L
AU  - Schomburg L
AD  - Max-Planck-Institut fur Experimentelle Endokrinologie, Hannover, Germany.
      lutz-schomburg@mail.uni-wuerzburg.de
FAU - Turwitt, S
AU  - Turwitt S
FAU - Prescher, G
AU  - Prescher G
FAU - Lohmann, D
AU  - Lohmann D
FAU - Horsthemke, B
AU  - Horsthemke B
FAU - Bauer, K
AU  - Bauer K
LA  - eng
SI  - GENBANK/AF126372
PT  - Comparative Study
PT  - Journal Article
PL  - England
TA  - Eur J Biochem
JT  - European journal of biochemistry
JID - 0107600
RN  - 0 (DNA, Complementary)
RN  - 0 (Recombinant Proteins)
RN  - 5Y5F15120W (Thyrotropin-Releasing Hormone)
RN  - EC 3.4.11.- (Aminopeptidases)
RN  - EC 3.4.19.6 (pyroglutamyl-peptidase II)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - SZB83O1W42 (Pyrrolidonecarboxylic Acid)
SB  - IM
MH  - Amino Acid Sequence
MH  - Aminopeptidases/*genetics/isolation & purification/metabolism
MH  - Chromosome Mapping
MH  - Chromosomes, Human, Pair 12
MH  - DNA, Complementary/genetics
MH  - Gene Library
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Kinetics
MH  - Metalloendopeptidases/*genetics/isolation & purification/metabolism
MH  - Molecular Sequence Data
MH  - Pyrrolidonecarboxylic Acid/analogs & derivatives
MH  - Recombinant Proteins/isolation & purification/metabolism
MH  - Sequence Analysis, DNA
MH  - Sequence Homology, Amino Acid
MH  - Species Specificity
MH  - Thyrotropin-Releasing Hormone/*metabolism
EDAT- 1999/09/22 00:00
MHDA- 1999/09/22 00:01
CRDT- 1999/09/22 00:00
PHST- 1999/09/22 00:00 [pubmed]
PHST- 1999/09/22 00:01 [medline]
PHST- 1999/09/22 00:00 [entrez]
AID - ejb753 [pii]
AID - 10.1046/j.1432-1327.1999.00753.x [doi]
PST - ppublish
SO  - Eur J Biochem. 1999 Oct 1;265(1):415-22. doi: 10.1046/j.1432-1327.1999.00753.x.