The Major Chemical-detoxifying System of UDP-glucuronosyltransferases Requires Regulated Phosphorylation Supported by Protein Kinase C

Basu, Nikhil K.; Kole, Labanyamoy; Basu, Mousumi; Chakraborty, Kushal; Mitra, Partha; Owens, Ida S.

doi:10.1074/jbc.m800032200

Cited by 29 publications

(67 citation statements)

References 50 publications

(85 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The His tag affinity ligand from the pcDNA3.1 vector (Invitrogen) was fused inframe at the 3Ј-end of pSVL-UGT2B15 cDNA (21,(22)(23)(24)(25). Clones were expressed in COS-1 and Src/Yes/Fyn (SYF ؊/؊ ) cells (American Type Culture Collection, Manassas, VA).…”

Section: Methodsmentioning

confidence: 99%

“…Clones were expressed in COS-1 and Src/Yes/Fyn (SYF ؊/؊ ) cells (American Type Culture Collection, Manassas, VA). Sources for all reagents used in tissue culture studies and the BCA protein assay kit were obtained as described (25). UGT substrates, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide kit, phorbol 12-myristate 13-acetate (PMA), anti-␤-actin antibody, 1,25-dihydroxyvitamin D 3 , PP2, and kinase inhibitors (curcumin, calphostin C, bisindolylmaleimide, Gö 6976, and röttlerin) were from either Sigma or Calbiochem.…”

Section: Methodsmentioning

confidence: 99%

“…Anti-His antibody was from BIOSOURCE (Camarillo, CA). The development of the highly specific anti-UGT-1168 antibody has been described (21,25). UDP-[…”

Section: Methodsmentioning

confidence: 99%

“…Glucuronidation Assay-Cellular extracts were analyzed for glucuronidation as described (25). The common donor substrate UDP-[…”

Section: Growth Transfection and Treatment Of Cells-cos-1 Monkey Kimentioning

confidence: 99%

“…Because ongoing in vitro studies (21)(22)(23)(24)(25), as well as evidence from an in vivo UGT model (26), have demonstrated each UGT requires regulated phosphorylation carried out by a different kinase, we sought to determine whether UGT2B15 is also dependent upon phosphorylation given that it has five phosphorylation sites: three PKC and two tyrosine kinase (TK) sites. Studies concerning the regulation of prostate-distributed UGTs that hasten DHT excretion from the body take on great importance because these preventive studies have indicated that elevated DHT is associated with prostate cancer and BPH development (11)(12)(13).…”

mentioning

confidence: 99%

See 4 more Smart Citations

Protein Kinase Cα and Src Kinase Support Human Prostate-distributed Dihydrotestosterone-metabolizing UDP-glucuronosyltransferase 2B15 Activity

Chakraborty¹,

Basu²,

Jana

et al. 2012

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Background: UDP-glucuronosyltransferase 2B15 (UGT2B15) is the only 5␣-dihydrotestosterone (DHT)-metabolizing enzyme in prostate luminal cells. Results: UGT2B15 requires regulated phosphorylation to metabolize DHT. Conclusion: PKC␣ and Src support phosphorylation of UGT2B15 at five sites involving two signaling pathways and cross-talk. Significance: Strategically controlled DHT synthesis and complex UGT2B15 phosphorylation impose low DHT turnover and presumably homeostatic levels of DHT-occupied androgen receptor for prostate-specific functions.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Anti-His antibody was from BIOSOURCE (Camarillo, CA). The development of the highly specific anti-UGT-1168 antibody has been described (21,25). UDP-[…”

Section: Methodsmentioning

confidence: 99%

“…Glucuronidation Assay-Cellular extracts were analyzed for glucuronidation as described (25). The common donor substrate UDP-[…”

Section: Growth Transfection and Treatment Of Cells-cos-1 Monkey Kimentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Protein Kinase Cα and Src Kinase Support Human Prostate-distributed Dihydrotestosterone-metabolizing UDP-glucuronosyltransferase 2B15 Activity

Chakraborty¹,

Basu²,

Jana

et al. 2012

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

Regulation of uridine diphosphate‐glucuronosyltransferase 1A3 activity by protein phosphorylation

Xiao

Yao

Jiang

et al. 2015

Biopharm & Drug Disp

View full text Add to dashboard Cite

Protein phosphorylation is a vital post-translational modification. This study investigated the effect of phosphorylation on human uridine diphosphate (UDP)-glucuronosyltransferase 1A3 (UGT1A3) activity. Curcumin and calphostin C suppressed the activity and phosphorylation of recombinant UGT1A3 expressed in Sf9 cells. These results indicate that UGT1A3 undergoes phosphorylation, which is required for its catalytic activity. Calphostin C is a highly specific protein kinase C (PKC) inhibitor, so three predicted PKC phosphorylation sites in UGT1A3 were examined. Site-directed mutation analysis at residues 28, 43 and 436 (from serine to glycine) was conducted. Compared with the wild-type, the S43G-mutant showed significantly decreased UGT1A3 catalytic activity. Furthermore, the UGT1A3 activity of wild-type and S43G-mutant was down-regulated by calphostin C, whereas the calphostin C inhibitory effect was much weaker on the S43G-mutant than the wild-type. In conclusion, phosphorylation plays an important role in UGT1A3 activity, and the serine at site 43 in UGT1A3 is most likely a phosphorylation site.

show abstract

Selective reduction in the expression of UGTs and SULTs, a novel mechanism by which piperine enhances the bioavailability of curcumin in rat

Zeng

Cai

Zeng

et al. 2017

Biopharm & Drug Disp

View full text Add to dashboard Cite

Curcumin (CUR) is known to exert numerous health-promoting effects in pharmacological studies, but its low bioavailability hinders the development of curcumin as a feasible therapeutic agent. Piperine (PIP) has been reported to enhance the bioavailability of curcumin, but the underlying mechanism remains poorly understood. In an attempt to find the mechanism by which piperine enhances the bioavailability of curcumin, the dosage ratio (CUR: PIP) and pre-treatment with piperine were hypothesized as key factors for improving the bioavailability in this combination. Therefore, combining curcumin with piperine at various dose ratios (1:1 to 100:1) and pre-dosing with piperine (0.5-8 h prior to curcumin) were designed to investigate their contributions to the pharmacokinetic parameters of curcumin in rats and their effects on the expression of UGT and SULT isoforms. It was shown that the C and AUC of curcumin were slightly increased by 1.29 and 1.67 fold at a ratio of 20:1, while curcumin exposure was enhanced significantly in all the piperine pre-treated rats (0.5-8 h), peaking at 6 h (a 6.09-fold and 5.97-fold increase in C and AUC , p < 0.01), regardless of the unchanged t and T . Also observed was a time-dependent inhibition of the hepatic expression of UGT1A6, 1A8, SULT1A1, 1A3, and the colonic expression of UGT1A6 that occurred within 6 h of piperine pre-treatment but was reversed at 8 h, which correlated with the changes in curcumin exposure. Similarly, the inhibitory effect of piperine on most of the UGTs and SULTs are time-dependent in Caco-2 and HepG2 cells. It is concluded that piperine pre-treatment time-dependently improves the bioavailability of curcumin through the reversible and selective inhibition of UGTs and SULTs. Copyright © 2016 John Wiley & Sons, Ltd.

show abstract

The Major Chemical-detoxifying System of UDP-glucuronosyltransferases Requires Regulated Phosphorylation Supported by Protein Kinase C

Cited by 29 publications

References 50 publications

Protein Kinase Cα and Src Kinase Support Human Prostate-distributed Dihydrotestosterone-metabolizing UDP-glucuronosyltransferase 2B15 Activity

Protein Kinase Cα and Src Kinase Support Human Prostate-distributed Dihydrotestosterone-metabolizing UDP-glucuronosyltransferase 2B15 Activity

Regulation of uridine diphosphate‐glucuronosyltransferase 1A3 activity by protein phosphorylation

Selective reduction in the expression of UGTs and SULTs, a novel mechanism by which piperine enhances the bioavailability of curcumin in rat

Contact Info

Product

Resources

About