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Isolation of a rat histidase cDNA sequence and expression in Escherichia coli--evidence of extrahepatic/epidermal distribution. Eur J Biochem 1997 Nov 15;250(1):212-21

Date

02/07/1998

Pubmed ID

9432011

DOI

10.1111/j.1432-1033.1997.00212.x

Scopus ID

2-s2.0-9844256437   11 Citations

Abstract

Histidase (histidine ammonia-lyase) is a cytosolic enzyme responsible for catalyzing the non-oxidative deamination of histidine to urocanic acid. Full-length cDNAs encoding rat histidase have been isolated from a lambdaZAP liver cDNA library using a partial cDNA fragment obtained by PCR. Whereas the initial description of the rat histidase 3' untranslated sequence contained a rare polyadenylation signal sequence, the data presented encompass a more distant 28-bp region, possessing a nucleotide stretch (AATATAAA), identical to that in the mouse histidase cDNA. Dideoxynucleotide chain-termination sequencing of two clones obtained by in vivo excision yielded an additional 376 bp and 105 bp of 5' and 3' untranslated sequences, respectively. A selected rat histidase cDNA clone was introduced into the pET-16b prokaryotic vector and expressed in BL21(DE3)pLysS Escherichia coli. After purification by nickel-chelation chromatography, recombinant histidine-tagged protein was employed to raise anti-(rat histidase) immunoglobulin in a Japanese white rabbit. The polyclonal rabbit antibody recognized and formed immune complexes with rat and recombinant human histidase proteins. Immunoblots of crude rat organ extracts detected a spectrum of histidase expression extending beyond that observed in liver and skin. Among other histidase-positive cells were those of the renal cortex tubular epithelium, fundic mucosal glands of stomach, gastric intramuscular (Auerbach's) plexus, and adrenal cortex. Immunohistochemical studies of histidase in rat liver produced discrete staining of hepatocytes in association with portal triads (Rappaport zone I). Furthermore, in contrast with previous reports of activity confined to epidermal stratum corneum, our findings demonstrate immunoreactive protein within and limited to the adjacent stratum granulosum.

Author List

Sano H, Tada T, Moriyama A, Ogawa H, Asai K, Kawai Y, Hodgson ME, Kato T, Wada Y, Suchi M

Author

Mariko Suchi MD, PhD Associate Professor in the Pathology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Animals
Antibodies
Base Sequence
Blotting, Western
Chromatography, Affinity
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel
Enzyme Inhibitors
Escherichia coli
Female
Histidine Ammonia-Lyase
Humans
Immunohistochemistry
Liver
Mice
Molecular Sequence Data
Rabbits
Rats
Recombinant Proteins
Sequence Analysis, DNA
Tissue Distribution
jenkins-FCD Prod-482 91ad8a360b6da540234915ea01ff80e38bfdb40a