Analysis and Modification of Thiols in the Hemolymph Juvenile Hormone Binding Protein of Manduca sexta

Two disulphide bridges are present in juvenile hormone binding protein from Galleria mellonella.

“…This strongly suggests that the two disulphide bridges reported previously for M. sexta JHBP should appear at Cys 9,16,151,195 . Since this protein alignment has shown that Cys 9,16,151,194 in B. mori [3] and Cys 9,16,150,194 in H. virescens [4] are conserved, therefore it is reasonable to postulate that JHBPs from these species also contain two disulphide bridges, which may be required for stabilizing the protein fold. It remains to be elucidated what is the topology at the -S-S bridge formation.…”

“…Comparison of amino-acid sequences of JHBPs from G. mellonella (Cys 10,17,151,195 ) (AF4107772), B. mori (Cys 9,16,151,194,200 ) [3], H. virescens (Cys 9,16,29,150,194 ) [4] and M. sexta (Cys 9,16,29,151,195,201 ) [5] shows that all Cys residues present in G. mellonella are conserved in the alignment of the proteins mentioned above (J.M. Rodriguez Parkitna et al, unpublished results).…”

“…It has been found that despite the substantial homology to the above men-tioned JHBPs orthologs this protein possesses only four Cys residues. The previous reports indicated that a JHBP molecule from M. sexta [9] contains two disulphide bridges and two free cysteine residues, therefore it was interesting to find if it is an obligatory feature of other JHBPs to have disulphide bridges and free cysteine residues in the molecule. An analysis of cystine and cysteine residues in G. mellonella JHBP is presented in this communication.…”

Evidences for a stage-specific juvenile hormone binding protein in the hemolymph of the silkworm,Bombyx mori L.: Identification and characterization by photoaffinity labeling and immunological analyses

“…In another case, Goodman et al (1990) observed multiple forms of JHBP in the hemolymph of Manduca sexta using two-dimensional Western blot and suggested that these cross-reacting proteins were either precursors or degradation products of native JHBP. In the recent study, Park and Goodman (1993) identified multiple electromorphs upon melanization of the hemolymph. They concluded that these multiple forms of JHBP were not the metabolites of JHBP since reducing SDS-PAGE yielded a single form of JHBP.…”

Purification and Characterization of Acid Phosphatase from the Egg of the Lady Beetle, Harmonia axyridis (Coccinellidae: Coleoptera)

“…Polyclonal antibodies made were used for detecting AP by Western blot. The procedures of the blotting (Towbin et al, 1979), was slightly modified (Park and Goodman, 1993). Briefly here, nitrocellulose paper (NCP, 0.2 µm, BIO-RAD) was used for the transfer of proteins after PAGE.…”