Positions of disulfide bonds and N-glycosylation site in juvenile hormone binding protein.
The juvenile hormone binding protein (JHBP) from Galleria mellonella hemolymph is a glycoprotein composed of 225 amino acid residues. It contains four Cys residues forming two disulfide bridges. In this study, the topography of the disulfide bonds as well as the site of glycan attachment in the JHBP molecule from G. mellonella was determined, using electrospray mass spectrometry. The MS analysis was performed on tryptic digests of JHBP. Our results show that the disulfide bridges link Cys10 and Cys17, and Cys151 and Cys195. Of the two potential N-glycosylation sites in JHBP, Asn4, and Asn94, only Asn94 is glycosylated. This site of glycosylation is also found in the fully biologically active recombinant JHBP expressed in the yeast Pichia pastoris.
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Poland.
Janusz
J
Debski
Aleksandra
A
Wysłouch-Cieszyńska
Michał
M
Dadlez
Krystyna
K
Grzelak
Barbara
B
Kłudkiewicz
Robert
R
Kołodziejczyk
Anna
A
Lalik
Andrzej
A
Ozyhar
Marian
M
Kochman
D000818:Animals; D002352:Carrier Proteins / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D003553:Cystine / Q000378:metabolism*; D006031:Glycosylation; D019476:Insect Proteins; D007814:Larva / Q000737:chemistry / Q000378:metabolism; D007915:Lepidoptera / Q000737:chemistry / Q000378:metabolism; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D020539:Sequence Analysis, Protein; D021241:Spectrometry, Mass, Electrospray Ionization
D002352:Carrier Proteins; D019476:Insect Proteins; D020033:Protein Isoforms; C035606:juvenile hormone-binding protein, insect; D003553:Cystine