Title:Finding Appropriate Signal Peptides for Secretory Production of Recombinant Glucarpidase: An In Silico Method
Volume: 15
Issue: 4
Author(s): Omid Vakili, Seyyed Hossein Khatami, Amir Maleksabet, Ahmad Movahedpour, Saeed Ebrahimi Fana, Rasoul Sadegh, Amir Hossein Salmanzadeh, Hadi Razeghifam, Sajjad Nourdideh, Sadra Samavarchi Tehrani*Mortaza Taheri-Anganeh*
Affiliation:
- Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran,Iran
- Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz,Iran
Keywords:
Methotrexate, recombinant protein, biopharmaceuticals, glucarpidase, bioinformatics, malignancies.
Abstract:
Background: Methotrexate (MTX) is a general chemotherapeutic agent utilized to treat
a variety of malignancies, woefully, its high doses can cause nephrotoxicity and subsequent defect
in the process of MTX excretion. The recombinant form of glucarpidase is produced by engineered
E. coli and is a confirmed choice to overcoming this problem.
Objective: In the present study, in silico analyses were performed to select suitable SPs for the secretion
of recombinant glucarpidase in E. coli.
Methods: The signal peptide website and UniProt database were employed to collect the SPs and
protein sequences. In the next step, SignalP-5.0 helped us to predict the SPs and the position of
cleavage sites. Moreover, physicochemical properties and solubility were evaluated using Prot-
Param and Protein-sol online software, and finally, ProtCompB was used to predict the final subcellular
localization.
Results: Luckily, all SPs could form soluble fusion proteins. At last, it was found that PPB and TIBA
could translocate the glucarpidase into the extracellular compartment.
Conclusion: This study showed that there are only 2 applicable SPs for the extracellular translocation
of glucarpidase. Although the findings were remarkable with high degrees of accuracy and precision
based on the utilization of bioinformatics analyses, additional experimental assessments are
required to confirm and validate it. Recent patents revealed several inventions related to the clinical
aspects of vaccine peptides against human disorders.
