PMID- 9178432
OWN - NLM
STAT- MEDLINE
DCOM- 19970811
LR  - 20190822
IS  - 0039-128X (Print)
IS  - 0039-128X (Linking)
VI  - 62
IP  - 5
DP  - 1997 May
TI  - The sheep kidney contains a novel unidirectional, high affinity NADP(+)-dependent 
      11 beta-hydroxysteroid dehydrogenase (11 beta-HSD-3).
PG  - 444-50
AB  - The 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) enzymes convert 
      corticosterone and cortisol to 11-dehydrocorticosterone and cortisone, and are 
      thought to convey extrinsic specificity to the mineralocorticoid receptor by 
      limiting access of the relatively more abundant glucocorticoids to it. Two 
      different 11 beta-hydroxysteroid dehydrogenases (11 beta-HSD) have been described 
      and cloned. The liver-type, NADP(+)-dependent 11 beta-HSD-1, has an affinity in 
      the micromolar range and bidirectional activity. The NAD(+)-dependent 11 
      beta-HSD-2 has a higher affinity, in the nanomolar range, and exhibits only 
      oxidase activity. 11 beta-HSD-2, because of its affinity and co-localization with 
      the mineralocorticoid receptor, is likely to serve as the "gatekeeper" for the 
      mineralocorticoid receptor in the kidney. Although the rat kidney expresses both 
      isoforms, only the high-affinity, NAD(+)-dependent 11 beta-HSD-2 has been 
      reported in the sheep kidney. We found both 11 beta-HSD NAD(+)- and 
      NADP(+)-dependent activities in sheep kidney to be present. The NAD(+)-dependent 
      activity exhibited a Km similar to that reported in the literature, 3.85 +/- 1.28 
      nM for corticosterone and 21.3 +/- 5.8 for cortisol, was distributed in 
      approximately equal amounts between microsomes and nuclei, and was 
      unidirectional, converting corticosterone to 11-dehydrocorticosterone. The enzyme 
      exhibited prominent substrate inhibition. The NADP(+)-dependent activity had a Km 
      for corticosterone of 4 +/- 1.3 nM for a Km for cortisol of 35.2 +/- 2 nM, 
      100-fold lower than that described for the 11 beta-HSD-1 in the liver of sheep 
      and other species, and was more prevalent in the microsomes than the nuclei. This 
      enzyme was not inhibited by its substrate. The NAD(+)-dependent activity was 
      approximately 3-10 times greater than the NADP(+)-dependent activity when 
      incubated with 5 nM corticosterone substrate, but had similar activity when 
      incubated with 100 nM substrate concentrations. CHOP cells (a modified Chinese 
      hamster ovary cell line) transiently transfected with the sheep 11 beta-HSD-2 
      plasmid exhibited a marked preference for NAD+ as co-factor. Oxidation of 
      corticosterone by transfected cells in the presence of NADP+ was present, but 
      minimal; NADP+ did not support the metabolism of cortisol, the primary 
      glucocorticoid of sheep. These data suggest the existence of another 
      NADP(+)-dependent enzyme, 11 beta-HSD-3, which, because of its high affinity and 
      unidirectional oxidase activity, may play a physiological role in the modulation 
      of glucocorticoid binding to both the mineralocorticoid and glucocorticoid 
      receptors.
FAU - Gomez-Sanchez, E P
AU  - Gomez-Sanchez EP
AD  - Research Service, Harry S. Truman Memorial Veterans Hospital, Columbia, MO 65201, 
      USA.
FAU - Ganjam, V
AU  - Ganjam V
FAU - Chen, Y J
AU  - Chen YJ
FAU - Cox, D L
AU  - Cox DL
FAU - Zhou, M Y
AU  - Zhou MY
FAU - Thanigaraj, S
AU  - Thanigaraj S
FAU - Gomez-Sanchez, C E
AU  - Gomez-Sanchez CE
LA  - eng
GR  - HL27255/HL/NHLBI NIH HHS/United States
GR  - HL27737/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Steroids
JT  - Steroids
JID - 0404536
RN  - 0U46U6E8UK (NAD)
RN  - 53-59-8 (NADP)
RN  - EC 1.1.- (Hydroxysteroid Dehydrogenases)
RN  - EC 1.1.1.146 (11-beta-Hydroxysteroid Dehydrogenases)
RN  - P540XA09DR (Glycyrrhetinic Acid)
RN  - W980KJ009P (Corticosterone)
SB  - IM
MH  - 11-beta-Hydroxysteroid Dehydrogenases
MH  - Animals
MH  - Corticosterone/metabolism
MH  - Glycyrrhetinic Acid/pharmacology
MH  - Hydroxysteroid Dehydrogenases/metabolism
MH  - Kidney/*enzymology
MH  - NAD/pharmacology
MH  - NADP/*pharmacology
MH  - Sheep
EDAT- 1997/05/01 00:00
MHDA- 1997/05/01 00:01
CRDT- 1997/05/01 00:00
PHST- 1997/05/01 00:00 [pubmed]
PHST- 1997/05/01 00:01 [medline]
PHST- 1997/05/01 00:00 [entrez]
AID - S0039-128X(97)00011-1 [pii]
AID - 10.1016/s0039-128x(97)00011-1 [doi]
PST - ppublish
SO  - Steroids. 1997 May;62(5):444-50. doi: 10.1016/s0039-128x(97)00011-1.