Identification of a renal-specific oxido-reductase in newborn diabetic mice

Qiwei Yang, Bharat Dixit, Jun Wada, Yufeng Tian, Elisabeth I. Wallner, Satish K. Srivastva, Yashpal S. Kanwar

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Aldose reductase (ALR2), a NADPH-dependent aldo-keto reductase (AKR), is widely distributed in mammalian tissues and has been implicated in complications of diabetes, including diabetic nephropathy. To identify a renal-specific reductase belonging to the AKR family, representational difference analyses of cDNA from diabetic mouse kidney were performed. A full-length cDNA with an ORF of 855 nt and yielding a ≃ 1.5-kb mRNA transcript was isolated from a mouse kidney library. Human and rat homologues also were isolated, and they had ≃ 91% and ≃ 97% amino acid identity with mouse protein. In vitro translation of the cDNA yielded a protein product of ≃ 33 kDa. Northern and Western blot analyses, using the cDNA and antirecombinant protein antibody, revealed its expression exclusively confined to the kidney. Like ALR2, the expression was up-regulated in diabetic kidneys. Its mRNA and protein expression was restricted to renal proximal tubules. The gene neither codistributed with Tamm-Horsfall protein nor aquaporin-2. The deduced protein sequence revealed an AKR-3 motif located near the N terminus, unlike the other AKR family members where it is confined to the C terminus. Fluorescence quenching and reactive blue agarose chromatography studies revealed that it binds to NADPH with high affinity (K(dNADPH) = 66.9 ± 2.3 nM). This binding domain is a tetrapeptide (Met-Ala-Lys-Ser) located within the AKR-3 motif that is similar to the other AKR members. The identified protein is designated as RSOR because it is renal-specific with properties of an oxido-reductase, and like ALR2 it may be relevant in the renal complications of diabetes mellitus.

Original languageEnglish (US)
Pages (from-to)9896-9901
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number18
DOIs
StatePublished - Aug 29 2000

Fingerprint

Oxidoreductases
Kidney
Complementary DNA
Proteins
Diabetes Complications
NADP
Uromodulin
Aquaporin 2
Aldehyde Reductase
Agarose Chromatography
Proximal Kidney Tubule
Messenger RNA
Diabetic Nephropathies
Northern Blotting
Open Reading Frames
carbonyl reductase (NADPH)
Fluorescence
Western Blotting
Amino Acids
Antibodies

Keywords

  • Diabetes mellitus
  • Diabetic nephropathy

ASJC Scopus subject areas

  • General

Cite this

Identification of a renal-specific oxido-reductase in newborn diabetic mice. / Yang, Qiwei; Dixit, Bharat; Wada, Jun; Tian, Yufeng; Wallner, Elisabeth I.; Srivastva, Satish K.; Kanwar, Yashpal S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 18, 29.08.2000, p. 9896-9901.

Research output: Contribution to journalArticle

Yang, Qiwei ; Dixit, Bharat ; Wada, Jun ; Tian, Yufeng ; Wallner, Elisabeth I. ; Srivastva, Satish K. ; Kanwar, Yashpal S. / Identification of a renal-specific oxido-reductase in newborn diabetic mice. In: Proceedings of the National Academy of Sciences of the United States of America. 2000 ; Vol. 97, No. 18. pp. 9896-9901.
@article{491acb8042844f5e9a69c966d81220b4,
title = "Identification of a renal-specific oxido-reductase in newborn diabetic mice",
abstract = "Aldose reductase (ALR2), a NADPH-dependent aldo-keto reductase (AKR), is widely distributed in mammalian tissues and has been implicated in complications of diabetes, including diabetic nephropathy. To identify a renal-specific reductase belonging to the AKR family, representational difference analyses of cDNA from diabetic mouse kidney were performed. A full-length cDNA with an ORF of 855 nt and yielding a ≃ 1.5-kb mRNA transcript was isolated from a mouse kidney library. Human and rat homologues also were isolated, and they had ≃ 91{\%} and ≃ 97{\%} amino acid identity with mouse protein. In vitro translation of the cDNA yielded a protein product of ≃ 33 kDa. Northern and Western blot analyses, using the cDNA and antirecombinant protein antibody, revealed its expression exclusively confined to the kidney. Like ALR2, the expression was up-regulated in diabetic kidneys. Its mRNA and protein expression was restricted to renal proximal tubules. The gene neither codistributed with Tamm-Horsfall protein nor aquaporin-2. The deduced protein sequence revealed an AKR-3 motif located near the N terminus, unlike the other AKR family members where it is confined to the C terminus. Fluorescence quenching and reactive blue agarose chromatography studies revealed that it binds to NADPH with high affinity (K(dNADPH) = 66.9 ± 2.3 nM). This binding domain is a tetrapeptide (Met-Ala-Lys-Ser) located within the AKR-3 motif that is similar to the other AKR members. The identified protein is designated as RSOR because it is renal-specific with properties of an oxido-reductase, and like ALR2 it may be relevant in the renal complications of diabetes mellitus.",
keywords = "Diabetes mellitus, Diabetic nephropathy",
author = "Qiwei Yang and Bharat Dixit and Jun Wada and Yufeng Tian and Wallner, {Elisabeth I.} and Srivastva, {Satish K.} and Kanwar, {Yashpal S.}",
year = "2000",
month = "8",
day = "29",
doi = "10.1073/pnas.160266197",
language = "English (US)",
volume = "97",
pages = "9896--9901",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "18",

}

TY - JOUR

T1 - Identification of a renal-specific oxido-reductase in newborn diabetic mice

AU - Yang, Qiwei

AU - Dixit, Bharat

AU - Wada, Jun

AU - Tian, Yufeng

AU - Wallner, Elisabeth I.

AU - Srivastva, Satish K.

AU - Kanwar, Yashpal S.

PY - 2000/8/29

Y1 - 2000/8/29

N2 - Aldose reductase (ALR2), a NADPH-dependent aldo-keto reductase (AKR), is widely distributed in mammalian tissues and has been implicated in complications of diabetes, including diabetic nephropathy. To identify a renal-specific reductase belonging to the AKR family, representational difference analyses of cDNA from diabetic mouse kidney were performed. A full-length cDNA with an ORF of 855 nt and yielding a ≃ 1.5-kb mRNA transcript was isolated from a mouse kidney library. Human and rat homologues also were isolated, and they had ≃ 91% and ≃ 97% amino acid identity with mouse protein. In vitro translation of the cDNA yielded a protein product of ≃ 33 kDa. Northern and Western blot analyses, using the cDNA and antirecombinant protein antibody, revealed its expression exclusively confined to the kidney. Like ALR2, the expression was up-regulated in diabetic kidneys. Its mRNA and protein expression was restricted to renal proximal tubules. The gene neither codistributed with Tamm-Horsfall protein nor aquaporin-2. The deduced protein sequence revealed an AKR-3 motif located near the N terminus, unlike the other AKR family members where it is confined to the C terminus. Fluorescence quenching and reactive blue agarose chromatography studies revealed that it binds to NADPH with high affinity (K(dNADPH) = 66.9 ± 2.3 nM). This binding domain is a tetrapeptide (Met-Ala-Lys-Ser) located within the AKR-3 motif that is similar to the other AKR members. The identified protein is designated as RSOR because it is renal-specific with properties of an oxido-reductase, and like ALR2 it may be relevant in the renal complications of diabetes mellitus.

AB - Aldose reductase (ALR2), a NADPH-dependent aldo-keto reductase (AKR), is widely distributed in mammalian tissues and has been implicated in complications of diabetes, including diabetic nephropathy. To identify a renal-specific reductase belonging to the AKR family, representational difference analyses of cDNA from diabetic mouse kidney were performed. A full-length cDNA with an ORF of 855 nt and yielding a ≃ 1.5-kb mRNA transcript was isolated from a mouse kidney library. Human and rat homologues also were isolated, and they had ≃ 91% and ≃ 97% amino acid identity with mouse protein. In vitro translation of the cDNA yielded a protein product of ≃ 33 kDa. Northern and Western blot analyses, using the cDNA and antirecombinant protein antibody, revealed its expression exclusively confined to the kidney. Like ALR2, the expression was up-regulated in diabetic kidneys. Its mRNA and protein expression was restricted to renal proximal tubules. The gene neither codistributed with Tamm-Horsfall protein nor aquaporin-2. The deduced protein sequence revealed an AKR-3 motif located near the N terminus, unlike the other AKR family members where it is confined to the C terminus. Fluorescence quenching and reactive blue agarose chromatography studies revealed that it binds to NADPH with high affinity (K(dNADPH) = 66.9 ± 2.3 nM). This binding domain is a tetrapeptide (Met-Ala-Lys-Ser) located within the AKR-3 motif that is similar to the other AKR members. The identified protein is designated as RSOR because it is renal-specific with properties of an oxido-reductase, and like ALR2 it may be relevant in the renal complications of diabetes mellitus.

KW - Diabetes mellitus

KW - Diabetic nephropathy

UR - http://www.scopus.com/inward/record.url?scp=0034730130&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034730130&partnerID=8YFLogxK

U2 - 10.1073/pnas.160266197

DO - 10.1073/pnas.160266197

M3 - Article

VL - 97

SP - 9896

EP - 9901

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 18

ER -