NDLI: Roles of Aromatic Residues in the Structure and Biological Activity of the Small Cytokine, Growth-blocking Peptide (GBP)

Content Provider	Journal of Biological Chemistry (JBC)
Author	Tada, Masahito Aizawa, Tomoyasu Shinohara, Yoshinori Matsubara, Kimiaki Miura, Kazunori Yoshida, Masanobu Shitara, Kunio Kouno, Takahide Mizuguchi, Mineyuki Nitta, Katsutoshi Hayakawa, Yoichi Kawano, Keiichi
Abstract	Growth-blocking peptide (GBP) is a small (25 amino acids) insect cytokine with a variety of functions: controlling the larval development of lepidopteran insects, acting as a mitogen for various types of cultured cells, and stimulating insect blood cells. The aromatic residues of GBP (Phe-3, Tyr-11, and Phe-23) are highly conserved in the ENF peptide family found in lepidopteran insects. We investigated the relationship between the biological activities and structural properties of a series of GBP mutants, in which each of the three aromatic residues is replaced by a different residue. The results of the hemocytes-stimulating assays of GBP mutants indicated that Phe-3 is the key residue in this activity: Ala or Tyr replacement resulted in significant loss of the activity, but Leu replacement did not. The replacements of other aromatic residues hardly affected the activity. On the other hand, NMR analysis of the mutants suggested that Tyr-11 is a key residue for maintaining the core structure of GBP. Surprisingly, the Y11A mutant maintained its biological activity, although its native-like secondary structure was disordered. Detailed analyses of the 15N-labeled Y11A mutant by heteronuclear NMR spectroscopy showed that the native-like β-sheet structure of Y11A was induced by the addition of 2,2,2-trifluoroethanol. The results suggest that Y11A has a tendency to form a native-like structure, and this property may give the Y11A mutant native-like activity.
Related Links	http://www.jbc.org/content/278/12/10778.abstract
Ending Page	10783
Starting Page	10778
Page Count	6
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	12
Volume Number	278
DOI	10.1074/jbc.M211251200
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2003-03-21
Access Restriction	Open
Subject Keyword	Growth-blocking peptide (GBP) Epidermal growth factor (EGF) High performance liquid chromatography (HPLC) Modified Grace's medium (MGM) Double quantum filtered correlation spectroscopy (DQF-COSY) Total correlation spectroscopy (TOCSY) Nuclear Overhauser effect spectroscopy (NOESY) Heteronuclear single quantum coherence (HSQC) Nuclear Overhauser effect (NOE) 2,2,2-trifluoroethanol (TFE) PROTEIN STRUCTURE AND FOLDING
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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