5 S,15 S-Dihydroperoxyeicosatetraenoic Acid (5,15-diHpETE) as a Lipoxin Intermediate: Reactivity and Kinetics with Human Leukocyte 5-Lipoxygenase, Platelet 12-Lipoxygenase, and Reticulocyte 15-Lipoxygenase-1.
5 S,15 S-Dihydroperoxyeicosatetraenoic Acid (5,15-diHpETE) as a Lipoxin Intermediate: Reactivity and Kinetics with Human Leukocyte 5-Lipoxygenase, Platelet 12-Lipoxygenase, and Reticulocyte 15-Lipoxygenase-1.
复制标题
DOI：
10.1021/acs.biochem.8b00889
复制
发表时间：
2018-12-04
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：
Green AR;Freedman C;Tena J;Tourdot BE;Liu B;Holinstat M;Holman TR
研究了5，15-diHpETE与人5-脂氧合酶(LOX)、人血小板12-LOX和网织红细胞15-LOX-1的反应活性和产生脂素(LXs)的相对速率。5-LOX不与5，15-diHpETE反应，但当15-HpETE为底物时，5-LOX能生成LXA4。相反，12-LOX和15-LOX-1都与5，15-diHpETE反应，专门形成LXB4。对于12-LOX和5，15-diHpETE，其动力学参数分别为kcat=0.17 S-1和kcat/Km=0.011μM-1s-1(分别比12-LOX氧化AA低106倍和1600倍)。另一方面，对于15-LOX-1，等效参数为kcat=4.6HpETE−1和kcat/Km=0.21HpETE M-1s-1(分别比AA的15-LOX-1地层的kCAT=4.6HpETE和Kcat/Km=0.21HpETE M-1s-1高3倍和相似)。这与15-LOX-2与5，15-diHpETE完全缺乏反应形成了鲜明对比(生物化学55,2832-2840,2016)。我们的数据表明，在催化5，15-diHpETE合成LXB4方面，12-LOX明显不如15-LOX-1。加入5，15-diHpETE可抑制血小板聚集，IC_(50)为1.3μM，而LXB_4在10μM以下对胶原介导的血小板活化无明显抑制作用。LXA4是含有5-LOX的主要产物，但必须以15-HpETE为底物。LXA4和LXB4的比例大致相同，由12-LOX产生，但只有在LTA4为底物的情况下，如前所述(Biochimica et BiPhysica Acta 1133,223-234,1992)。
The reaction of 5,15-diHpETE with human 5-lipoxygenase (LOX), human platelet 12-LOX and human reticulocyte 15-LOX-1 was investigated to determine the reactivity and relative rates of producing lipoxins (LXs). 5-LOX does not react with 5,15-diHpETE, although it can produce LXA4 when 15-HpETE is the substrate. In contrast, both 12-LOX and 15-LOX-1 react with 5,15-diHpETE, forming specifically LXB4. For 12-LOX and 5,15-diHpETE, the kinetic parameters are kcat = 0.17 s-1 and kcat/KM = 0.011 μM-1s-1 (106-fold and 1600-fold lower than for 12-LOX oxygenation of AA, respectively). On the other hand, for 15-LOX-1 the equivalent parameters are kcat = 4.6 s−1 and kcat/KM = 0.21 μM-1s-1 (3-fold higher and similar to that for 12-HpETE formation by 15-LOX-1 from AA, respectively). This contrasts with the complete lack of reaction of 15-LOX-2 with 5,15-diHpETE (Biochemistry 55, 2832–2840, 2016). Our data indicate that 12-LOX is markedly inferior to 15-LOX-1 in catalyzing the production of LXB4 from 5,15-diHpETE. Platelet aggregation was inhibited by the addition of 5,15-diHpETE, with an IC50 of 1.3 μM, however, LXB4 did not significantly inhibit collagen-mediated platelet activation up to 10 μM. In summary, LXB4 is the primary product of 12-LOX and 15-LOX-1 catalysis if 5,15-diHpETE is the substrate, with 15-LOX-1 being 20-fold more efficient than 12-LOX. LXA4 is the primary product with 5-LOX, but only if 15-HpETE is the substrate. Approximately equal proportions of LXA4 and LXB4 are produced by 12-LOX, but only if LTA4 is the substrate, as described previously (Biochimica et Biophysica Acta 1133, 223–234, 1992).
登录
查看更多内容
DOI：
10.1016/0005-2760(93)90085-n
发表时间：
1993-07-21
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
KUHN, H;BARNETT, J;SIGAL, E
通讯作者：
SIGAL, E
影响因子：
2.9
作者：
Segraves, EN;Holman, TR
通讯作者：
Holman, TR
影响因子：
2.9
作者：
Green, Abigail R.;Barbour, Shannon;Holman, Theodore R.
通讯作者：
Holman, Theodore R.
DOI：
10.1016/0167-4889(92)90073-k
发表时间：
1992-01-13
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
SHEPPARD, KA;GREENBERG, SM;SERHAN, CN
通讯作者：
SERHAN, CN
DOI：
10.1016/0006-291x(84)91486-4
发表时间：
1984-01-01
影响因子：
3.1
作者：
SERHAN, CN;HAMBERG, M;SAMUELSSON, B
通讯作者：
SAMUELSSON, B