Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Nov;295(5):F1535-44.
doi: 10.1152/ajprenal.90445.2008. Epub 2008 Sep 10.

Coenzyme Q10 supplementation rescues renal disease in Pdss2kd/kd mice with mutations in prenyl diphosphate synthase subunit 2

Ryoichi Saiki  1 , Adam L Lunceford, Yuchen Shi, Beth Marbois, Rhonda King, Justin Pachuski, Makoto Kawamukai, David L Gasser, Catherine F Clarke
Affiliations

Coenzyme Q10 supplementation rescues renal disease in Pdss2kd/kd mice with mutations in prenyl diphosphate synthase subunit 2

Ryoichi Saiki et al. Am J Physiol Renal Physiol. 2008 Nov.

Abstract

Homozygous mice carrying kd (kidney disease) mutations in the gene encoding prenyl diphosphate synthase subunit 2 (Pdss2kd/kd) develop interstitial nephritis and eventually die from end-stage renal disease. The PDSS2 polypeptide in concert with PDSS1 synthesizes the polyisoprenyl tail of coenzyme Q (Q or ubiquinone), a lipid quinone required for mitochondrial respiratory electron transport. We have shown that a deficiency in Q content is evident in Pdss2kd/kd mouse kidney lipid extracts by 40 days of age and thus precedes the onset of proteinuria and kidney disease by several weeks. The presence of the kd (V117M) mutation in PDSS2 does not prevent its association with PDSS1. However, heterologous expression of the kd mutant form of PDSS2 together with PDSS1 in Escherichia coli recapitulates the Q deficiency observed in the Pdss2kd/kd mouse. Dietary supplementation with Q10 provides a dramatic rescue of both proteinuria and interstitial nephritis in the Pdss2kd/kd mutant mice. The results presented suggest that Q may be acting as a potent lipid-soluble antioxidant, rather than by boosting kidney mitochondrial respiration. Such Q10 supplementation may have profound and beneficial effects in treatment of certain forms of focal segmental glomerulosclerosis that mirror the renal disease of the Pdss2kd/kd mouse.

PubMed Disclaimer

Figures

Fig. 1.
Fig. 1.
Amino acid sequence alignment of mouse nonaprenyl diphosphate synthase subunit 1 (PDSS1/mSPS1) and subunit 2 (PDSS2/mDLP1) isoforms. A: typical long-chain polyisoprenyl diphosphate synthases, such as PDSS1 and PDSS2-L, have conserved domains I through VII. An arrow shows the location of the kd/kd mutation V117M within conserved domain I. B: alternative splicing produces PDSS2-Short (PDSS2-S) containing exons 1–3b and PDSS2-Long (PDSS2-L) containing exons 1–3a and 4–8. PDSS2-S contains only domains I through III. The predicted polypeptide produced from the Pdss2 BAC clone 256E1 is also shown. One transgenic line of B6.kd/kd mice harboring an insertion of this BAC clone 256E1 exhibits sporadic rescue from kidney disease.
Fig. 2.
Fig. 2.
Kidney Q9 and Q10 levels are significantly lower in B6.Pdss2kd/kd mice than in B6 wild-type mice. Wild-type B6, heterozygous B6.+/kd, and mutant B6.Pdss2kd/kd mice were killed at the ages shown, and Q levels were measured in kidney homogenates prepared from each mouse. Error bars indicate the standard deviation in 3 separate measurements of the same lipid extract. Levels of Q9 and Q10 in 123- and 151-day-old B6.Pdss2kd/kd mice and in 115- and 149-day-old B6 mice were previously reported (33).
Fig. 3.
Fig. 3.
Differences in liver Q9 content in B6.Pdss2kd/kd mice and B6 wild-type mice are evident by 149 days of age. Wild-type B6 and mutant B6.Pdss2kd/kd mice were killed at the ages shown, and Q levels were measured in liver homogenates prepared from each mouse. Error bars indicate the standard deviation in 3 separate measurements of the same lipid extract.
Fig. 4.
Fig. 4.
Expression of a truncated form of Pdss2 in B6.Pdss2kd/kd mice restores content of Q in kidney extracts to wild-type levels in most cases. Wild-type B6, mutant B6.Pdss2kd/kd, and line G B6.Pdss2kd/kd transgenic mice (indicated by asterisks) were killed at the ages shown, and Q9 and Q10 levels were measured in kidney homogenates taken from each mouse. Error bars indicate the standard deviation in 3 separate measurements of the same lipid extract.
Fig. 5.
Fig. 5.
Heterologous expression of the mutant kd form of Pdss2 in Escherichia coli recapitulates the deficiency in Q biosynthesis observed in B6.Pdss2kd/kd mice. A: pGEX-KG (empty vector) produces just the glutathione S-transferase (GST) polypeptide. pM12S contains the open reading frame (ORF) of Pdss1 inserted in frame after the carboxyl terminus of GST (GST-PDSS1) and Pdss2-S (Short form) inserted in frame after a six-Histidine tag (His6-PDSS2-S). pM12L expresses GST-PDSS1 and His6-PDSS2-L (Long form). pM12Lkd expresses GST-PDSS1 and His6-PDSS2-L-V117M (Long form harboring the kd mutation). B, BamHI; E, EcoRI; S, SalI, and Xb, XbaI. An asterisk shows the kd/kd mutation position V117M, and “rbs” indicates a ribosomal binding site from pET-28c to allow polycistronic expression of the mouse polypeptides. B: Q8, normally produced in E. coli, and Q9, generated by the heterologous expression of PDSS1 and PDSS2, were measured in lipid extracts prepared from E. coli DH5α harboring each of the constructs designated in A. E. coli strain KO229 harboring a deletion of the ispB prenyl diphosphate synthase gene was rescued by pM12L. Error bars indicate the standard deviation in 3 separate measurements of the same lipid extract.
Fig. 6.
Fig. 6.
Coprecipitation of PDSS1 and PDSS2 polypeptides. His-PDSS2-L or His-PDSS2-L-V117M polypeptides were incubated with GST-PDSS1 for 30 min and subsequently precipitated by Ni-NTA Superflow resin (lanes 2 and 3). A coprecipitation assay of GST and His-PDSS2-L also was performed as a negative control at the same time (lane 1). Antibodies to GST (A) and the 6-His tag (B) were utilized to recognize GST-PDSS1 and His-PDSS2 polypeptides, respectively. The asterisks indicate nonspecific bands recognized by the GST antibody. GST*(truncated), observed in lanes 2, 3, 5, and 6 in A, bottom, is likely to result from proteolysis of the GST-PDSS1 fusion protein. Lanes 4, 5, and 6 show the relative amounts of input protein used for each coprecipitation assay and correspond to lanes 1, 2, and 3, respectively. Similar amounts of GST-PDSS1 were coprecipitated with both His-PDSS2-L and His-PDSS2-Lkd fusion proteins (A, lanes 2 and 3, top). In contrast, GST did not coprecipitate with the His-PDSS1-L fusion protein (A, lane 1, bottom).
Fig. 7.
Fig. 7.
Western blot analysis of PDSS2 polypeptides in mice liver mitochondria. An anti-PDSS2 antibody was utilized to detect PDSS2 polypeptides in 120 μg of mouse mitochondria protein (A). An existing amount of PDSS2-L and -S in B6 mouse mitochondria (lane 1) was significantly higher than in kd/kd and alb/cre mutant mice (lanes 2 and 3, respectively). PDSS2-S is present in relatively smaller amounts than PDSS2-L. An anti-yeast F1βATP synthase subunit that cross-reacts with mouse F1βATP synthase subunit provided a loading control (B).
Fig. 8.
Fig. 8.
Hematoxylin- and eosin-stained sections of kidneys from Q10-treated and untreated B6.Pdss2kd/kd mice. A: no histological evidence of disease in a 162-day-old female treated with Q10. The amount of urine albumin measured was 3.7 mg/24 h. B: severe interstitial nephritis in an untreated 128-day-old female. The amount of urine albumin measured was 57.34 mg/24 h. Note the severe interstitial inflammation, tubular dilatation, and glomerular crescents. Magnification, ×100.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Backhed F, Manchester JK, Semenkovich CF, Gordon JI. Mechanisms underlying the resistance to diet-induced obesity in germ-free mice. Proc Natl Acad Sci USA 104: 979–984, 2007. - PMC - PubMed
    1. Barisoni L, Diomedi-Camassei F, Santorelli FM, Caridi G, Thomas DB, Emma F, Piemonte F, Ghiggeri GM. Collapsing glomerulopathy associated with inherited mitochondrial injury. Kidney Int 74: 237–243, 2008. - PubMed
    1. Barisoni L, Madaio MP, Eraso M, Gasser DL, Nelson PJ. The kd/kd mouse is a model of collapsing glomerulopathy. J Am Soc Nephrol 16: 2847–2851, 2005. - PMC - PubMed
    1. Bentinger M, Brismar K, Dallner G. The antioxidant role of coenzyme Q. Mitochondrion 7S: S41–S50, 2007. - PubMed
    1. Bishop NA, Guarente L. Genetic links between diet and lifespan: shared mechanisms from yeast to humans. Nat Rev Genet 8: 835–844, 2007. - PubMed

Publication types

MeSH terms