داکینگ مولکولی پپتید باکتریوسین انتروسین P با آنتی‌ژن سطحی باکتری اشرشیاکلای مولد ورم پستان در گاوشیری

نوع مقاله : مقاله کامل

نویسندگان

گروه علوم دامی، دانشکده کشاورزی، دانشگاه فردوسی مشهد

چکیده

استفاده بیش‌ازحد از آنتی‌بیوتیک‌ها برای درمان بیماری‌هایی مانند ورم پستان گاوهای شیری ناشی از اشرشیاکلای، منجر به ظهور باکتری‌های مقاوم شده است که تهدیدی برای سلامت عمومی قلمداد می‌شود. امروزه پپتیدهای ضدمیکروبی نظیر باکتریوسین‌ها به‌عنوان عوامل بالقوه ضدمیکروبی توجه بیشتری به خود جلب کرده‌اند؛ بنابراین بررسی پایداری و فعالیت ضدباکتریایی آن‌ها در شرایط بدن حیوان بسیار حائز اهمیت می‌باشد. لذا، باهدف بررسی پایداری حرارتی پپتید انتروسین P ن (P (EntP، ابتدا ساختار سوم EntP با استفاده از سرور I-TASSER پیش‌بینی شد. سپس، پایداری این ساختار در شرایط دینامیکی مولکولی مورد ارزیابی قرار گرفت. بدین منظور شبیه‌سازی با کمک نرم‌افزار GROMACS در مدت ‌زمان‌ ns 400 برای دمای طبیعی بدن گاوشیری (K°312) صورت گرفت. درنهایت منحنی RMSD مربوط به این پپتید در مقایسه با زمان، در طول خط سیر شبیه‌سازی رسم گردید. به‌منظور بررسی برهم‌کنش پپتید با پروتئین لیپوپلی‌ساکارید سطحی D (LptD از باکتری اشرشیاکلای مولد ورم پستان از سرور آنلاین ClusPro استفاده شد. در بخش آزمایشگاهی نیز حداقل غلظت مهارکنندگی (MIC) و حداقل غلظت کشندگی (MBC) براساس روش میکروبراث بر روی اشریشیاکلای برآورد گردید. براساس نتایج گزارش‌شده توسط سرور I-TASSER، بهترین ساختار پیش‌بینی‌شده‌ی پپتید EntP با ضریب اطمینان (C-score) برابر با 4/1 انتخاب شد. مقادیر گزارش‌شده توسط پلات‌های ERRAT و Verify3D نشان داد که ساختار سوم پیش‌بینی‌شده برای پپتید EntP رضایت‌بخش است. گراف RMSD مربوط به بررسی پایداری حرارتی EntP در دمای بدن گاو نشان داد که ساختار پپتید EntP پس از مدت ns 400 دارای خط سیر شبیه‌سازی پایدار است. نتایج مربوط به داکینگ مولکولی نیز نشان داد این پپتید به آنتی‌ژن LptD در موقعیت مناسب متصل می‌شود. میزان حداقل غلظت بازدارندگی (MIC) و حداقل غلظت کشندگی (MBC) به ترتیب 25/48 و5/96 (μg/ml) برای باکتری اشرشیاکلای برآورد شدند. امید است در آینده بتوان از این پپتید به‌عنوان ترکیبی جایگزین آنتی‌بیوتیک یا در کنار آن به‌منظور درمان بیماری‌های دامی با منشا اشرشیاکلای استفاده کرد.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular docking of bacteriocin enterocin P peptide with mastitis-causing E. coli antigen in cattle

نویسندگان [English]

  • Zahra Mousavi
  • Zahra Rashidian
  • Yasaman Zeraatpisheh
  • A. Javadmanesh
Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
چکیده [English]

Excessive use of antibiotics to treat diseases such as mastitis in dairy cows caused by Escherichia coli has led to the emergence of antibiotic-resistant bacteria, which is considered a threat to public health. Nowadays, antimicrobial peptides such as bacteriocins have attracted more attention as potential antimicrobial agents. Therefore, the study of their stability and antibacterial activity in physiological conditions is very important. In order to investigate the thermal stability of Enterocin P peptide (EntP), the third structure of EntP was first predicted using the I-TASSER server. Then, the stability of this structure was evaluated by dynamic conditions for this purpose, the simulation was performed using GROMACS software in 400 ns at cow body temperature (312 °K). Finally, the root-mean-square deviation (RMSD) graph of this peptide was plotted against time along the simulation trajectory. ClusPro online server was used to study the interaction of peptide with LptD surface protein from Escherichia coli bacterium causing mastitis. In the in vitro phase of this study, the minimum inhibitory concentration (MIC) and minimum dissociation concentration (MBC) of EntP peptide were estimated based on the microbroth method on Escherichia coli. The results showed that the best predicted structure of EntP peptide by I-TASSER server had a C-score of 1.4 which was selected. The values reported by the ERRAT and Verify3D plots indicated that the tertiary structure predicted for the EntP peptide was satisfactory. The RMSD graph related to the thermal stability of EntP at bovine body temperature showed that the structure of EntP peptide is stable after 400 ns of the simulation trajectory. The results of molecular docking also showed that this peptide was correctly attached to the LptD antigen. After measuring the MIC, the bacteria were cultured and the MBC was obtained, which were estimated at 48.25 and 96.5 μg/ml, respectively. This peptide could be used as an alternative to antibiotics or in combination with them for the treatment of infections with the origin of Escherichia coli.

کلیدواژه‌ها [English]

  • EntP
  • Peptide
  • Bioinformatics
  • Mastitis
  • Molecular docking
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