The study of the mechanism of the action between the positive/variable natural molecules and the 5-lipooxygenase.

Guide: It is assumed that AKBA can promote the formation of SPM in 5-LOX-rich neutrophils, mononucleoblasts, or macrophages by inducing the transition of 5-LOX region specificity to 12/15-lipoylyoxetase5-lipoxygenase (5-lipoxygenase, 5-LOX) is a dioxyase that contains non-hemoglobin ions in an organism and is a key enzyme that catalyzes the production of leukotriene (leukotrienes, LTs) of peanut tetraoleic acid (arachidonic acid, AA)LTs are an important inflammatory medium and participate in many diseases such as atherosclerosis, cardiovascular disease, heart disease and other pathological physiological processesIn addition, the study found that 5-LOX was over-expression in a variety of cancer cells, such as prostate, oral, colon, and bladder cancerAs a result, 5-LOX is considered an important drug target for a variety of diseases, and researchers have been working on 5-LOX inhibitorsRecently, researchers at Louisiana State University in the United States explored the different mechanisms of action between two natural products, the redox inhibitor NDGA at the active site and the variant inhibitor AKBA and 5-LOXfirst,, they analyzed the crystal structure of the resolution of the 2.71-5-LOX and NDGA complexes, and found that the inhibitor NDGA induced the remodeling of the catalytic region of 5-LOX, exposed the NDGA to the deep valleys of the active site, and the inhibitor NDGA only built a bridge between the valleys by interacting with Arg596The structural comparison analysis shows that the composite structure is completely different from the stable apo5-LOX closed structureFurther analysis found that the ndGA's two phthalaterings occupied a certain spatial position, resulting in the anchoring of pyridoxene Phe177, Tyr 181, Phe421 and Asn425 moving outward, thus causing the disorder of the active regionThe researchers then studied the sensitivity of 5-LOX to protein hydrolysis when they bind to different substratesThe study found that 5-LOX's sensitivity to protein hydrolysis did not change when 5-LOX interacted with AA, but when NDGA existed, 5-LOX's sensitivity to protein hydrolysis increased 5 times, confirming that inhibitor NDGA could cause disorder in the structure of the 5-LOX active site regionat the same time, the researchers analyzed the crystal structure of the resolution of the 3.0-5-LOX and AKBA complexes and found that AKBA was located in a deep groove between the amino end and the catalytic region, with a distorted chair structure at one end of the argon and acetic acid base, which interacted with Arg101, Thr137, Arg138 and Val110The other end is provided by Leu66, Val110, Ile126 and Lys133Structural analysis showed that combined with AKBA, the highly conservative cation-to-thoracity interaction between the amino end Ofamine Trp102 and the catalytic region Arg165, and the salt bridge between Arg101 and Asp166 disappeared, indicating that the signaling effect between the regions that can be mediated was destroyedCombined with Bfactors analysis, it is further shown that the combination of the variable-structure inhibitor AKBA can affect interregional mobility and changes in 5-LOX conformationlater, the researchers analyzed how the inhibitors NDGA and AKBA regulate 5-LOX activity by monitoring the formation of the classic enzyme products 5-HETE and LTB4 of peanut tetraoleic AA The experimental results show that the inhibition of NDGA and AKBA to 5-LOX is increased in concentration dependence In addition, the researchers monitored the production of by-products 12-HETE and 15-HETE AKBA resulted in a significant increase in the production of 12-HETE, of which 10 s.M was most pronounced, but 15-HETE was not affected In contrast, NDGA is concentrated on a dependency that inhibits the production of 12-HETE and 15-HETE Taking into account the complex regulation of 5-LOX in the cellular environment, the researchers monitored the inhibition effect of NDGA and AKBA on 5-LOX in HE293 cells by ultra-efficient liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to further evaluate the effects of the inhibitory effects of the inhibitors of the inhibitors OF NDGA and AKBA in the cell environment In HEK293 cells, NDGA is effective in suppressing the production of all 5-LOX products AKBA significantly reduces the production of 5-LOX products, especially LTB4 and its isomers At the same time, the researchers found a significant increase in by-product 12-HETE in the cellular environment In addition, the researchers demonstrated that AKBA's regulatory effect was only effective at 5-LOX, and they simultaneously expressed 12-LOX, 15-LOX-1 and 15-LOX-2 in HEK239 cells The experimental results show that AKBA has no inhibitory effect on other LOXs These data show that AKBA induction causes a change in the individuality of the 5-LOX region, which leads to the production of by-product 12-HETE, and is only effective for cells expressing 5-LOX, indicating that AKBA is isomer-specific to the transformational regulation of 5-LOX in addition, the researchers confirmed by expressing 5-LOX in human neutrophils and macrophages MDM that AKBA-induced production of 12-HETE occurs under associated pathophysiological conditions Because SPM synthesis requires 12/15-LOX activity, the researchers speculate that AKBA can promote the formation of SPM in 5-LOX-rich neutrophils, mononucleosis, or macrophages by inducing the 5-LOX region to transition to 12/15-lipoycozyne In summary, the variant inhibitor AKBA not only inhibits the formation of 5-LOX classic enzyme products, but also causes a change in the specificity of the 5-LOX region This study provides a new strategy for the development of isomer-specific morphoid inhibitors at 5-LOX's variable-structure sites reference Gilbert, N.C., Gerstmeier, J., Schexnaydre, E.E Etal Rhandiai and mechanistic insights into 5-lipoxygenase inhibition bynatural products Nat Chem Biol (2020) .