Roles of aromatic residues in the structure and biological activity of the small cytokine, growth-blocking peptide (GBP)

被引:8
|
作者
Tada, M
Aizawa, T
Shinohara, Y
Matsubara, K
Miura, K
Yoshida, M
Shitara, K
Kouno, T
Mizuguchi, M
Nitta, K
Hayakawa, Y
Kawano, K
机构
[1] Toyama Med & Pharmaceut Univ, Fac Pharmaceut Sci, Toyama 9300194, Japan
[2] Biooriented Technol Res Advancement Inst, Omiya, Saitama 3318537, Japan
[3] Hokkaido Univ, Grad Sch Neurosci, Div Biol Sci, Sapporo, Hokkaido 0600810, Japan
[4] Hokkaido Univ, Inst Low Temp Sci, Sapporo, Hokkaido 0600819, Japan
关键词
D O I
10.1074/jbc.M211251200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 N-15-labeled Y11A mutant by heteronuclear NMR spectroscopy showed that the native-like beta-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.
引用
收藏
页码:10778 / 10783
页数:6
相关论文
共 27 条
  • [1] Role of aromatic residues in the structure and biological activity of the small cytokine, growth-blocking peptide (GBP)
    Tade, M
    Aizawa, T
    Mizuguchi, M
    Nitta, K
    Hayakawa, Y
    Kawano, K
    BIOPOLYMERS, 2003, 71 (03) : 315 - 315
  • [2] Role of aromatic residues in the structure and biological activity of the small cytokine, growth-blocking peptide (GBP)
    Tada, Masahito
    Aizawa, Tomoyasu
    Mizuguchi, Mineyuki
    Nitta, Katsutoshi
    Hayakawa, Yoichi
    Kawano, Keiichi
    PEPTIDE REVOLUTION: GENOMICS, PROTEOMICS & THERAPEUTICS, 2004, : 468 - 469
  • [3] CONTRIBUTION OF FLEXIBLE N-TERMINAL RESIDUES TO THE BIOLOGICAL ACTIVITY OF INSECT CYTOKINE, GROWTH-BLOCKING PEPTIDE (GBP)
    Yoshida, M.
    Shitara, K.
    Matsubara, K.
    Kouno, T.
    Aizawa, T.
    Hayakawa, Y.
    Kumaki, Y.
    Mizuguchi, M.
    Demura, M.
    Nitta, K.
    Kawano, K.
    JOURNAL OF PEPTIDE SCIENCE, 2004, 10 : 226 - 226
  • [4] Structure and activity of the insect cytokine growth-blocking peptide
    Aizawa, T
    Hayakawa, Y
    Ohnishi, A
    Fujitani, N
    Clark, KD
    Strand, MR
    Miura, K
    Koganesawa, N
    Kumaki, Y
    Demura, M
    Nitta, K
    Kawano, K
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (34) : 31813 - 31818
  • [5] Insect cytokine growth-blocking peptide (GBP) regulates insect development
    Hayakawa, Yoichi
    APPLIED ENTOMOLOGY AND ZOOLOGY, 2006, 41 (04) : 545 - 554
  • [6] Cell growth activity of growth-blocking peptide
    Hayakawa, Y
    Ohnishi, A
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 250 (02) : 194 - 199
  • [7] Cloning of growth-blocking peptide (gbp)-inducible phosphorylated protein that is involved in hemocyte activation
    Oda, Yasunori
    Hayakawa, Yoichi
    ZOOLOGICAL SCIENCE, 2005, 22 (12) : 1497 - 1497
  • [8] The Gly-Gly linker region of the insect cytokine growth-blocking peptide is essential for activity
    Yoshida, M
    Aizawa, T
    Nakamura, T
    Shitara, K
    Hayakawa, Y
    Matsubara, K
    Miura, K
    Kouno, T
    Clark, KD
    Strand, MR
    Mizuguchi, M
    Demura, M
    Nitta, K
    Kawano, K
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (49) : 51331 - 51337
  • [9] Solution structure of an insect growth factor, growth-blocking peptide
    Aizawa, T
    Fujitani, N
    Hayakawa, Y
    Ohnishi, A
    Ohkubo, T
    Kumaki, Y
    Kawano, K
    Hikichi, K
    Nitta, K
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (04) : 1887 - 1890