Biosynthesis of the Maresin Intermediate, 13S,14S-Epoxy-DHA, by Human 15-Lipoxygenase and 12-Lipoxygenase and Its Regulation through Negative Allosteric Modulators.
Biosynthesis of the Maresin Intermediate, 13S,14S-Epoxy-DHA, by Human 15-Lipoxygenase and 12-Lipoxygenase and Its Regulation through Negative Allosteric Modulators.
复制标题
DOI：
10.1021/acs.biochem.0c00233
复制
发表时间：
2020-05-19
期刊：
影响因子：
2.9
通讯作者：
Holman TR
中科院分区：
文献类型：
--
作者：
Freedman C;Tran A;Tourdot BE;Kalyanaraman C;Perry S;Holinstat M;Jacobson MP;Holman TR
人网织红细胞15-脂氧合酶-1（h15-LOX-1或ALOX 15）和血小板12-脂氧合酶（h12-LOX或ALOX 12）催化二十二碳六烯酸（DHA）和maresin前体，14 S-氢过氧基-4Z，7Z，10 Z，12 E，16 Z，19 Z-二十二碳六烯酸（14 S-HpDHA），以确定其在关键maresin中间体13 S，14 S-epoxy-4 Z生物合成中的产物分布和相对速率，7Z，9 E，11 E，16 Z，19 Z-二十二碳六烯酸（13 S，14 S-环氧-DHA）。两种酶都能将DHA转化为14 S-HpDHA，其中h12-LOX的kcat/KM值（14.0 ± 0.8 s−1 μM−1）是h15-LOX-1的39倍（0.36 ± 0.08 s−1 μ M −1），14 S-HpDHA产物选择性是h15-LOX-1的1.8倍（81%和46%）。然而，h12-LOX在从14 S-HpDHA生产13 S，14 S-epoxy-DHA方面的效率明显低于h15-LOX-1，kcat/KM值分别为0.0024 ± 0.0002和0.11 ± 0.006 s−1 μM−1，是h15-LOX-1的4.6倍。这是h15-LOX-1催化该反应的第一个证据，并揭示了maresin生物合成的一种新的体外途径。此外，14 S-HpDHA的环氧化通过与h15-LOX-1和h12-LOX结合的变构氧脂素负调控。对于h15-LOX-1，14 S-HpDHA（Kd = 6.0 μM）、12 S-hydroxy-5 Z，8 Z，10 E，14 Z-二十碳四烯酸（12 S-HETE）（Kd = 3.5 μM）和14 S-hydroxy-7Z，10 Z，12 E，16 Z，19 Z-二十二碳五烯酸（14 S-HDPA ω-3）（Kd = 4.0 μM）显示降低13 S，14 S-epoxy-DHA的产生。h12-LOX也显示出受14 S-HpDHA（Kd = 3.5 μM）和14 S-HDPA ω-3（Kd = 4.0 μM）的变构调节;然而，12 S-HETE没有显示出任何影响，这首次表明h12-LOX的变构反应。最后，14 S-HpDHA在亚微摩尔浓度下抑制血小板聚集，这可能与富含DHA的饮食的益处有关。这些体外生物合成途径可能有助于指导体内maresin生物合成研究和可能的直接治疗干预。
Human reticulocyte 15-lipoxygenase-1 (h15-LOX-1 or ALOX15) and platelet 12-lipoxygenase (h12-LOX or ALOX12) catalysis of docosahexaenoic acid (DHA) and the maresin precursor, 14S-hydroperoxy-4Z,7Z,10Z,12E,16Z,19Z-docosahexaenoic acid (14S-HpDHA), were investigated to determine their product profiles and relative rates in the biosynthesis of the key maresin intermediate, 13S,14S-epoxy-4Z,7Z,9E,11E,16Z,19Z-docosahexaenoic acid (13S,14S-epoxy-DHA). Both enzymes converted DHA to 14S-HpDHA, with h12-LOX having a 39-fold greater kcat/KM value (14.0 ± 0.8 s−1 μM−1) than that of h15-LOX-1 (0.36 ± 0.08 s−1 μM−1) and a 1.8-fold greater 14S-HpDHA product selectivity, 81 and 46%, respectively. However, h12-LOX was markedly less effective at producing 13S,14S-epoxy-DHA from 14S-HpDHA than h15-LOX-1, with a 4.6-fold smaller kcat/KM value, 0.0024 ± 0.0002 and 0.11 ± 0.006 s−1 μM−1, respectively. This is the first evidence of h15-LOX-1 to catalyze this reaction and reveals a novel in vitro pathway for maresin biosynthesis. In addition, epoxidation of 14S-HpDHA is negatively regulated through allosteric oxylipin binding to h15-LOX-1 and h12-LOX. For h15-LOX-1, 14S-HpDHA (Kd = 6.0 μM), 12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid (12S-HETE) (Kd = 3.5 μM), and 14S-hydroxy-7Z,10Z,12E,16Z,19Z-docosapentaenoic acid (14S-HDPAω-3) (Kd = 4.0 μM) were shown to decrease 13S,14S-epoxy-DHA production. h12-LOX was also shown to be allosterically regulated by 14S-HpDHA (Kd = 3.5 μM) and 14S-HDPAω-3 (Kd = 4.0 μM); however, 12S-HETE showed no effect, indicating for the first time an allosteric response by h12-LOX. Finally, 14S-HpDHA inhibited platelet aggregation at a submicrololar concentration, which may have implications in the benefits of diets rich in DHA. These in vitro biosynthetic pathways may help guide in vivo maresin biosynthetic investigations and possibly direct therapeutic interventions.
登录
查看更多内容
影响因子：
4.8
作者：
Coffa, G;Imber, AN;Brash, AR
通讯作者：
Brash, AR
影响因子：
3.5
作者：
Ivanov I;Kuhn H;Heydeck D
通讯作者：
Heydeck D
DOI：
10.1161/atvbaha.117.309868
发表时间：
2017-10
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Adili R;Tourdot BE;Mast K;Yeung J;Freedman JC;Green A;Luci DK;Jadhav A;Simeonov A;Maloney DJ;Holman TR;Holinstat M
通讯作者：
Holinstat M
影响因子：
1.8
作者：
Rodriguez, Ana R.;Spur, Bernd W.
通讯作者：
Spur, Bernd W.
影响因子：
8.6
作者：
Pistorius K;Souza PR;De Matteis R;Austin-Williams S;Primdahl KG;Vik A;Mazzacuva F;Colas RA;Marques RM;Hansen TV;Dalli J
通讯作者：
Dalli J