Glifosatın üreaz enzimi üzerindeki interferansına ilişkin in vitro deneysel bir çalışma

Year 2024, Volume: 49 Issue: 3, 779 - 792, 30.09.2024

Kezban Kartlaşmış , Mehmet Tahir Hüsunet , Sevinç Püren Yücel Karakaya , Tuğba Polat , Zeynep Tan , Nurten Dikmen

https://doi.org/10.17826/cumj.1514902

Abstract

Amaç: Türkiye'de tarım alanlarının yoğunluğu nedeniyle glifosata maruziyet artmaktadır. Bu çalışmada glifosatın birçok hastalığın tanı ve takibinde üre ölçümünde sıklıkla kullanılan bir enzim olan üreaz üzerindeki olası interferans etkisi incelendi.
Gereç ve Yöntem: Ön denemelerde glifosatın, değişen konsantrasyonlarda üre solüsyonlarıyla negatif etkileşime sahip olduğu gözlendi. İn vitro ortamda glifosat eklenmesi öncesi ve sonrası sonuçlara etkisini belirlemek amacıyla kan örnekleri üreaz-GLDH ve dolaylı nesslerizasyon prosedürleri kullanılarak incelendi. Morfolojik ve kimyasal değişiklikler için taramalı elektron mikroskobu (SEM) ve Fouirer-Transform Infrared Spektrofotometre (FTIR) analizleri yapıldı ve moleküler kenetleme yoluyla bağlanma modelleri oluşturuldu. Üreaz-GLDH ve dolaylı nesslerizasyon ile gerçekleştirilen üre ölçümleri, 10–3, 10–4, ve 10–5 M konsantrasyonlarında glifosattaki sonuçlar üzerinde negatif bir etkileşim göstermiştir.
Bulgular: SEM analizinde gözlenen morfolojik değişiklikler, FTIR analizinde oluşan 3228.25 (O-H), 1642.08 (C=C), ve 1531.20 (N-O) cm–1 bağları ile desteklenmiştir. Ayrıca moleküler kenetleme analizi, glifosatın, üreazı hidrojen bağı (Gly13, Ser12, Lys14, Thr15, ve Asp37) ve hidrofobik etkileşimler (Val10, Asp37, ve Glu98) yoluyla etkilediğini göstermektedir. Bu etkileşimli amino asitlerin, üreazın aktif katalitik konformasyonunun erişilebilirliğini sınırladığını ve/veya katalitik geçiş durumunun stabilitesini etkilediği öngörülmüştür.
Sonuç: Glifosat, insan serum üre testlerinde negatif etkileşime yol açarak klinik biyokimya, mikrobiyoloji ve tarım laboratuvarlarında hatalı test sonuçlarına yol açar. Analiz yapılırken bu etki dikkate alınmalı ve klinisyenlerin yanı sıra hastane bilgi yönetim sistemleri de bu müdahaleye özel önem verilerek önceden bilgilendirilmelidir.

Keywords

Glifosat, girişim, üreaz

An in vitro experimental study on the interference of glyphosate on the urease enzyme

Abstract

Purpose: Exposure to glyphosate is increasing due to the density of agricultural areas in Türkiye. In this study, the possible interference effect of glyphosate on urease, an enzyme that is frequently used in the diagnosis and follow-up of many diseases and in the measurement of urea in biological samples was examined.
Materials and Methods: First, glyphosate was observed to have a negative interference in experiments using solutions of varying concentrations of urea. Second, blood samples were examined using the urease-glutamate dehydrogenase (GLDH) and indirect nesslerization procedures to determine the effects of glyphosate on the results before and after its addition. To determine the morphological and chemical alterations, scanning electron microscope (SEM) and Fourier-transform infrared spectroscopy (FTIR) analyses were conducted, and binding patterns were established through molecular docking. Urea measurements conducted with urease-GLDH and indirect nesslerization demonstrated a negative interference on the results with glyphosate concentrations of 10–3, 10–4, and 10–5 M.
Results: Morphological changes observed in the SEM analysis were supported by the 3228.25 (O-H), 1642.08 (C=C), and 1531.20 (N-O) cm–1 bonds formed in the FTIR analysis. Furthermore, the molecular docking analysis showed that glyphosate affected the urease via hydrogen bonding (Gly13, Ser12, Lys14, Thr15, and Asp37) and hydrophobic interactions (Val10, Asp37, and Glu98). It was hypothesized that these interacting amino acids limit the accessibility of the urease’s active catalytic conformation and/or impact the stability of the catalytic transition state.
Conclusion: Glyphosate leads to negative interference in human serum urea assays, leading to incorrect test results in clinical biochemistry, microbiology, and agricultural laboratories. This effect should be considered when conducting analysis, and clinicians as well as hospital information management systems should be informed ahead of time, with special emphasis devoted to this interference.

Keywords

glyphosate, urease, interference

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