PMID- 10620514
OWN - NLM
STAT- MEDLINE
DCOM- 20000302
LR  - 20181113
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 345 Pt 2
DP  - 2000 Jan 15
TI  - Molecular cloning, sequencing and expression studies of the human breast cancer
      cell glutaminase.
PG  - 365-75
AB  - Phosphate-activated glutaminase (GA) is overexpressed in certain types of tumour 
      but its exact role in tumour cell growth and proliferation is unknown. Here we
      describe the isolation of a full-length cDNA clone of human breast cancer ZR75
      cells, by a combination of lambdagt10 cDNA library screening and the rapid
      amplification of cDNA ends ('RACE') technique. The cDNA of human GA is 2408 nt
      with a 1806-base open reading frame encoding a 602-residue protein with a
      predicted molecular mass of 66309 Da. The deduced amino acid sequence contains a 
      putative mitochondrial import presequence of 14 residues at the N-terminal end.
      Heterologous expression and purification in Escherichia coli yielded a product of
      the expected molecular size that was recognized by using antibodies against the
      recombinant human GA. Sequence analyses showed that human GA was highly similar
      to the rat liver enzyme. Northern gel analysis revealed that the gene is present 
      in human liver, brain and pancreas, in which a major transcript of 2.4 kb was
      demonstrated, but not in kidney, heart, skeletal muscle, lung or placenta. These 
      results strongly suggest that the first human GA cloned, the GA from ZR-75 breast
      cancer cells, and presumably those from human liver and brain, are liver-type
      isoenzymes, in sharp contrast with the present view that considers the kidney
      type as the isoform expressed in all tissues with GA activity, with the exception
      of postnatal liver.
FAU - Gomez-Fabre, P M
AU  - Gomez-Fabre PM
AD  - Departamento de Biologia Molecular y Bioquimica, Laboratorio de Quimica de
      Proteinas, Facultad de Ciencias, Universidad de Malaga, Campus de Teatinos, 29071
      Malaga, Spain.
FAU - Aledo, J C
AU  - Aledo JC
FAU - Del Castillo-Olivares, A
AU  - Del Castillo-Olivares A
FAU - Alonso, F J
AU  - Alonso FJ
FAU - Nunez De Castro, I
AU  - Nunez De Castro I
FAU - Campos, J A
AU  - Campos JA
FAU - Marquez, J
AU  - Marquez J
LA  - eng
SI  - GENBANK/AF110330
SI  - GENBANK/AF110331
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0RH81L854J (Glutamine)
RN  - EC 3.5.1.2 (Glutaminase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Breast Neoplasms/*enzymology
MH  - Cloning, Molecular
MH  - Escherichia coli/genetics
MH  - Female
MH  - Gene Expression
MH  - Gene Library
MH  - Glutaminase/biosynthesis/*genetics
MH  - Glutamine/*metabolism
MH  - Humans
MH  - Molecular Sequence Data
MH  - Neoplasm Proteins/biosynthesis/*genetics
MH  - Recombinant Proteins/biosynthesis
MH  - Sequence Analysis, DNA
MH  - Sequence Homology, Amino Acid
MH  - Tissue Distribution
PMC - PMC1220766
EDAT- 2000/01/06 09:00
MHDA- 2000/03/04 09:00
CRDT- 2000/01/06 09:00
PHST- 2000/01/06 09:00 [pubmed]
PHST- 2000/03/04 09:00 [medline]
PHST- 2000/01/06 09:00 [entrez]
PST - ppublish
SO  - Biochem J. 2000 Jan 15;345 Pt 2:365-75.