Expression and Characterization of Manduca sexta Stress Responsive Peptide-1; An Inducer of Antimicrobial Peptide Synthesis
Biochemistry and Molecular Biology
Volume 4, Issue 3, May 2019, Pages: 42-52
Received: Jul. 5, 2019; Accepted: Aug. 1, 2019; Published: Aug. 14, 2019
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Lynn Goss Schrag, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, USA
Xiaolong Cao, Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, USA
Hawa Dembele, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, USA
Xiaorong Liu, Department of Chemistry, University of Massachusetts, Amherst, USA
Qasim Al Souhail, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, USA
Michael Robert Kanost, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, USA
Jianhan Chen, Department of Chemistry, University of Massachusetts, Amherst, USA; Department of Biohemistry and Molecular Biology, University of Massachusetts, Amherst, USA
Haobo Jiang, Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, USA
Om Prakash, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, USA
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In response to stress conditions such as wounding or infections in insects, several short peptides are processed to act as cytokines that induce AMP gene expression. To study their structure-activity relationship, immune inducibility, tissue specificity, stress responsiveness, and development relatedness, we chemically synthesized Manduca sexta stress response peptide-1, a 25-residue peptide with one disulfide bond (SRP1: FGVRVGTCPSGYVRRGTFCFPDDDY). Upon injection of the SRP1 into naïve larvae, several antimicrobial peptide genes were expressed at higher levels. The mRNA levels of SRP1 increased significantly in hemocytes and fat body after larvae were challenged with a mixture of bacteria and β-1,3-glucan. The expression patterns of SRP1 and its target genes are somewhat different from SRP2’s, suggesting overlapping yet distinct functions. We elucidated the 3D structure of SRP1 in solution by two-dimensional 1H-1H NMR spectroscopy. The tertiary structure of SRP1 consists of two short β-strands at Y12−R15 and F18−F20, one type-II β-turn at R15−F18 in its well-defined core and is stabilized by a covalent disulfide bond between C8 and C19. The conformational ensemble of SRP1 from extensive atomistic simulation in explicit solvent (with 3.0μs total effective sampling) shows high consistency with experimental intramolecular NOEs of the core region. The SRP1 core adopts a fold similar to the carboxyl-terminal subdomain of epidermal growth factor (EGF), suggesting that SRP1 may interact with EGF receptor-like molecules to trigger its biological function.
Cytokine, Nuclear Magnetic Resonance, Molecular Dynamics, Epidermal Growth Factor, Hemolymph Protein, Insect Immunity, Phenoloxidase
To cite this article
Lynn Goss Schrag, Xiaolong Cao, Hawa Dembele, Xiaorong Liu, Qasim Al Souhail, Michael Robert Kanost, Jianhan Chen, Haobo Jiang, Om Prakash, Expression and Characterization of Manduca sexta Stress Responsive Peptide-1; An Inducer of Antimicrobial Peptide Synthesis, Biochemistry and Molecular Biology. Vol. 4, No. 3, 2019, pp. 42-52. doi: 10.11648/j.bmb.20190403.12
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