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 -