UV-difference and CD spectroscopy studies on juvenile hormone binding to its carrier protein.
Juvenile hormone binding protein (JHBP) from hemolymph of Galleria mellonella is a single-chain glycoprotein of molecular mass near 25,880 containing no Trp residues. The fourth derivative of the protein absorption spectrum shows the characteristic vibrational components of the phenylalanine spectrum within the range 240-270 nm. At longer wavelengths two main bands 0-0 and 0+800 cm(-1) appear, caused by vibrational levels of electronic transition, pi-->pi*, in the tyrosine residues, with maxima at 279 nm and 286 nm, respectively. Two intersection points of the second derivative absorption band with abscissa at about 288.8 nm and 283 nm were analysed for estimation of the environment polarity of Tyr residues in the JHBP molecule. The results obtained suggest that JHBP contains at least two classes of Tyr residues with very apolar environment, similar to that found in azurine. In the JHBP-JH complex only one class of Tyr residues located in a very apolar environment was found, and a small perturbation of disulphide bridges was deduced from the UV-difference spectrum. Ligand perturbation appears as a minimum at 243 nm of the UV-difference spectrum. Comparison of the circular dichroism (CD) spectra for free JHBP with the CD spectra for the JHBP-JH complex monitored in the far-UV (190-240 nm) region indicates rather small differences in the secondary structure of the protein. Although ligand binding induces distinct changes in the near-UV (250-300 nm) region of the CD spectrum of JHBP, it is apparent where both Tyr and Phe residues contribute.
Krzyzanowska
D
D
Division of Biochemistry, Institute of Organic Chemistry, Biochemistry and Biotechnology, Technical University of Wroclaw, Poland.
Lisowski
M
M
Kochman
M
M
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D006458:Hemolymph / Q000737:chemistry; D019476:Insect Proteins; D007814:Larva; D009036:Moths; D010649:Phenylalanine / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D014443:Tyrosine / Q000737:chemistry
D002352:Carrier Proteins; D019476:Insect Proteins; D012717:Sesquiterpenes; C035606:juvenile hormone-binding protein, insect; D014443:Tyrosine; D010649:Phenylalanine; C036585:juvenile hormone III