Structure and Activity of the Insect Cytokine Growth-blocking Peptide

Growth-blocking peptide (GBP) is a 25-amino acid insect cytokine found in Lepidopteran insects that possesses diverse biological activities such as larval growth regulation, cell proliferation, and stimulation of immune cells (plasmatocytes). The tertiary structure of GBP consists of a structured core that contains a disulfide bridge and a short antiparallel β-sheet (Tyr11–Arg13 and Cys19–Pro21) and flexible N and C termini (Glu1–Gly6 and Phe23–Gln25). In this study, deletion and point mutation analogs of GBP were synthesized to investigate the relationship between the structure of GBP and its mitogenic and plasmatocyte spreading activity. The results indicated that deletion of the N-terminal residue, Glu1, eliminated all plasmatocyte spreading activity but did not reduce mitogenic activity. In contrast, deletion of Phe23 along with the remainder of the C terminus destroyed all mitogenic activity but only slightly reduced plasmatocyte spreading activity. Therefore, the minimal structure of GBP containing mitogenic activity is 2–23 GBP, whereas that with plasmatocyte spreading activity is 1–22 GBP. NMR analysis indicated that these N- and C-terminal deletion mutants retained a similar core structure to wild-type GBP. Replacement of Asp16 with either a Glu, Leu, or Asn residue similarly did not alter the core structure of GBP. However, these mutants had no mitogenic activity, although they retained about 50% of their plasmatocyte spreading activity. We conclude that specific residues in the unstructured and structured domains of GBP differentially affect the biological activities of GBP, which suggests the possibility that multifunctional properties of this peptide may be mediated by different forms of a GBP receptor.

[1]  B. Volkman,et al.  Alanine-scanning Mutagenesis of Plasmatocyte Spreading Peptide Identifies Critical Residues for Biological Activity* , 2001, The Journal of Biological Chemistry.

[2]  Clark,et al.  Plasmatocyte spreading peptide (PSP1) and growth blocking peptide (GBP) are multifunctional homologs. , 2000, Journal of insect physiology.

[3]  B. Sykes,et al.  NMR study of the differential contributions of residues of transforming growth factor alpha to association with its receptor. , 2000, Protein engineering.

[4]  M. Kanost,et al.  Structure of a paralytic peptide from an insect, Manduca sexta. , 2001, The journal of peptide research : official journal of the American Peptide Society.

[5]  M. Kanost,et al.  Biological activity of Manduca sexta paralytic and plasmatocyte spreading peptide and primary structure of its hemolymph precursor. , 1999, Insect biochemistry and molecular biology.

[6]  H. Kataoka,et al.  Isolation and structure determination of a paralytic peptide from the hemolymph of the silkworm, Bombyx mori , 1999, Peptides.

[7]  J. Markley,et al.  Structure of the Insect Cytokine Peptide Plasmatocyte-spreading Peptide 1 from Pseudoplusia includens * , 1999, The Journal of Biological Chemistry.

[8]  J. Shabanowitz,et al.  A cardioactive peptide from the southern armyworm, Spodoptera eridania , 1999, Peptides.

[9]  T. Aizawa,et al.  Solution Structure of an Insect Growth Factor, Growth-blocking Peptide* , 1999, The Journal of Biological Chemistry.

[10]  A. Ohnishi,et al.  Cell growth activity of growth-blocking peptide. , 1998, Biochemical and biophysical research communications.

[11]  M. Strand,et al.  Isolation and Identification of a Plasmatocyte-spreading Peptide from the Hemolymph of the Lepidopteran InsectPseudoplusia includens * , 1997, The Journal of Biological Chemistry.

[12]  Y. Hayakawa,et al.  Role of dopamine at the onset of pupal diapause in the cabbage armyworm Mamestra brassicae , 1997, FEBS letters.

[13]  D. Hoyt,et al.  NMR Study of the Transforming Growth Factor-α (TGF-α)-Epidermal Growth Factor Receptor Complex , 1996, The Journal of Biological Chemistry.

[14]  A. Ohnishi,et al.  Molecular cloning and characterization of cDNA for insect biogenic peptide, growth‐blocking peptide , 1995, FEBS letters.

[15]  A. Ohnishi,et al.  Growth-blocking peptide titer during larval development of parasitized and cold-stressed armyworm. , 1995, Insect biochemistry and molecular biology.

[16]  D. Davies,et al.  Contribution of the Transforming Growth Factor B-loop -Sheet to Binding and Activation of the Epidermal Growth Factor Receptor (*) , 1995, The Journal of Biological Chemistry.

[17]  A. Lenferink,et al.  Identification of the high affinity binding site of transforming growth factor-alpha (TGF-alpha) for the chicken epidermal growth factor (EGF) receptor using EGF/TGF-alpha chimeras. , 1994, The Journal of biological chemistry.

[18]  Y. Hayakawa Inclusion of a beta-aspartyl peptide bond causes the loss of inherent physiological activities. , 1994, Journal of biochemistry.

[19]  Y. Hayakawa,et al.  Growth-blocking peptide or polydnavirus effects on the last instar larvae of some insect species , 1993 .

[20]  G. B. Quistad,et al.  Paralytic peptides from hemolymph of the lepidopteran insect Trichoplusia ni Hubner. , 1993, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[21]  D. Davies,et al.  Multidomain binding of transforming growth factor alpha to the epidermal growth factor receptor. , 1992, Biochemistry.

[22]  J. Tam,et al.  Mapping the receptor-recognition site of human transforming growth factor-alpha. , 2009, International journal of peptide and protein research.

[23]  G. B. Quistad,et al.  Isolation and identification of paralytic peptides from hemolymph of the lepidopteran insects Manduca sexta, Spodoptera exigua, and Heliothis virescens. , 1991, The Journal of biological chemistry.

[24]  Y. Hayakawa Structure of a growth-blocking peptide present in parasitized insect hemolymph. , 1991, The Journal of biological chemistry.

[25]  G. Fields,et al.  Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids. , 2009, International journal of peptide and protein research.

[26]  Y. Hayakawa Juvenile hormone esterase activity repressive factor in the plasma of parasitized insect larvae. , 1990, The Journal of biological chemistry.

[27]  E. Pedroso,et al.  Formation of aspartimide peptides in Asp-Gly sequences , 1989 .

[28]  E. Olejniczak,et al.  Improvement of 2D NOE spectra of biomacromolecules in H2O solution by coherent suppression of the solvent resonance , 1986 .

[29]  R. B. Merrifield,et al.  A synthetic approach to structure-function relationships in the murine epidermal growth factor molecule. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[30]  K. Wüthrich,et al.  Improved spectral resolution in cosy 1H NMR spectra of proteins via double quantum filtering. , 1983, Biochemical and biophysical research communications.

[31]  Richard R. Ernst,et al.  Coherence transfer by isotropic mixing: Application to proton correlation spectroscopy , 1983 .

[32]  L. Hood,et al.  A gas-liquid solid phase peptide and protein sequenator. , 1981, The Journal of biological chemistry.

[33]  Dieter Suter,et al.  Two-dimensional chemical exchange and cross-relaxation spectroscopy of coupled nuclear spins , 1981 .