Abstract
vitamin D is 25-hydroxylated in the liver, before being activated by 1α-hydroxylation in the kidney. Recently, the rat cytochrome P450 2J3 (CYP2J3) has been identified as a principal vitamin D 25-hydroxylase in the rat [Yamasaki T, Izumi S, Ide H, Ohyama Y. Identification of a novel rat microsomal vitamin D3 25-hydroxylase. J Biol Chem 2004;279(22):22848-56]. In this study, we examine whether human CYP2J2 that exhibits 73% amino acid homology to rat CYP2J3 has similar catalytic properties. Recombinant human CYP2J2 was overexpressed in Escherichia coli, purified, and assayed for vitamin D 25-hydroxylation activity. We found significant 25-hydroxylation activity toward vitamin D3 (turnover number, 0.087 min-1), vitamin D2 (0.16 min-1), and 1α-hydroxyvitamin D3 (2.2 min-1). Interestingly, human CYP2J2 hydroxylated vitamin D2, an exogenous vitamin D, at a higher rate than it did vitamin D3, an endogenous vitamin D, whereas, rat CYP2J3 hydroxylated vitamin D3 (1.4 min-1) more efficiently than vitamin D2 (0.86 min-1). Our study demonstrated that human CYP2J2 exhibits 25-hydroxylation activity as well as rat CYP2J3, although the activity of human CYP2J2 is weaker than rat CYP2J3. CYP2J2 and CYP2J3 exhibit distinct preferences toward vitamin D3 and D2.
| Original language | English |
|---|---|
| Pages (from-to) | 849-856 |
| Number of pages | 8 |
| Journal | Steroids |
| Volume | 71 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - Oct 1 2006 |
| Externally published | Yes |
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Keywords
- 25-Hydroxylase
- CYP2J2
- CYP2J3
- vitamin D
- vitamin D
ASJC Scopus subject areas
- Biochemistry
- Endocrinology
- Molecular Biology
Cite this
Characterization of rat and human CYP2J enzymes as Vitamin D 25-hydroxylases. / Aiba, Isamu; Yamasaki, Tomoaki; Shinki, Toshimasa; Izumi, Shunsuke; Yamamoto, Keiko; Yamada, Sachiko; Terato, Hiroaki; Ide, Hiroshi; Ohyama, Yoshihiko.
In: Steroids, Vol. 71, No. 10, 01.10.2006, p. 849-856.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of rat and human CYP2J enzymes as Vitamin D 25-hydroxylases
AU - Aiba, Isamu
AU - Yamasaki, Tomoaki
AU - Shinki, Toshimasa
AU - Izumi, Shunsuke
AU - Yamamoto, Keiko
AU - Yamada, Sachiko
AU - Terato, Hiroaki
AU - Ide, Hiroshi
AU - Ohyama, Yoshihiko
PY - 2006/10/1
Y1 - 2006/10/1
N2 - vitamin D is 25-hydroxylated in the liver, before being activated by 1α-hydroxylation in the kidney. Recently, the rat cytochrome P450 2J3 (CYP2J3) has been identified as a principal vitamin D 25-hydroxylase in the rat [Yamasaki T, Izumi S, Ide H, Ohyama Y. Identification of a novel rat microsomal vitamin D3 25-hydroxylase. J Biol Chem 2004;279(22):22848-56]. In this study, we examine whether human CYP2J2 that exhibits 73% amino acid homology to rat CYP2J3 has similar catalytic properties. Recombinant human CYP2J2 was overexpressed in Escherichia coli, purified, and assayed for vitamin D 25-hydroxylation activity. We found significant 25-hydroxylation activity toward vitamin D3 (turnover number, 0.087 min-1), vitamin D2 (0.16 min-1), and 1α-hydroxyvitamin D3 (2.2 min-1). Interestingly, human CYP2J2 hydroxylated vitamin D2, an exogenous vitamin D, at a higher rate than it did vitamin D3, an endogenous vitamin D, whereas, rat CYP2J3 hydroxylated vitamin D3 (1.4 min-1) more efficiently than vitamin D2 (0.86 min-1). Our study demonstrated that human CYP2J2 exhibits 25-hydroxylation activity as well as rat CYP2J3, although the activity of human CYP2J2 is weaker than rat CYP2J3. CYP2J2 and CYP2J3 exhibit distinct preferences toward vitamin D3 and D2.
AB - vitamin D is 25-hydroxylated in the liver, before being activated by 1α-hydroxylation in the kidney. Recently, the rat cytochrome P450 2J3 (CYP2J3) has been identified as a principal vitamin D 25-hydroxylase in the rat [Yamasaki T, Izumi S, Ide H, Ohyama Y. Identification of a novel rat microsomal vitamin D3 25-hydroxylase. J Biol Chem 2004;279(22):22848-56]. In this study, we examine whether human CYP2J2 that exhibits 73% amino acid homology to rat CYP2J3 has similar catalytic properties. Recombinant human CYP2J2 was overexpressed in Escherichia coli, purified, and assayed for vitamin D 25-hydroxylation activity. We found significant 25-hydroxylation activity toward vitamin D3 (turnover number, 0.087 min-1), vitamin D2 (0.16 min-1), and 1α-hydroxyvitamin D3 (2.2 min-1). Interestingly, human CYP2J2 hydroxylated vitamin D2, an exogenous vitamin D, at a higher rate than it did vitamin D3, an endogenous vitamin D, whereas, rat CYP2J3 hydroxylated vitamin D3 (1.4 min-1) more efficiently than vitamin D2 (0.86 min-1). Our study demonstrated that human CYP2J2 exhibits 25-hydroxylation activity as well as rat CYP2J3, although the activity of human CYP2J2 is weaker than rat CYP2J3. CYP2J2 and CYP2J3 exhibit distinct preferences toward vitamin D3 and D2.
KW - 25-Hydroxylase
KW - CYP2J2
KW - CYP2J3
KW - vitamin D
KW - vitamin D
UR - http://www.scopus.com/inward/record.url?scp=33747863597&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33747863597&partnerID=8YFLogxK
U2 - 10.1016/j.steroids.2006.04.009
DO - 10.1016/j.steroids.2006.04.009
M3 - Article
C2 - 16842832
AN - SCOPUS:33747863597
VL - 71
SP - 849
EP - 856
JO - Steroids
JF - Steroids
SN - 0039-128X
IS - 10
ER -