Purification and initial characterization of a novel protein with factor Xa activity from Lonomia obliqua caterpillar spicules.
A novel protein with factor Xa-like activity was isolated from Lonomia obliqua caterpillar spicules by gel filtration chromatography and reversed-phase high-performance liquid chromatography. The protein had a mass of 20745.7 Da, as determined by mass spectrometry, and contained four Cys residues. Enzymatic hydrolysis followed by de novo sequencing by tandem mass spectrometry was used to determine the primary structure of the protein and the cysteine residues linked by disulfide bridges. The positions of 24 sequenced tryptic peptides, including the N-terminal, were deduced by comparison with a homologous protein from the superfamily Bombycoidea. Approximately 90% of the primary structure of the active protein was determined.
Lilla
S
S
Department of Pharmacology, Institute of Biomedical Sciences, University of São Paulo (USP), São Paulo, SP, Brazil.
Pereira
R
R
Hyslop
S
S
Donato
J L
JL
Le Bonniec
B F
BF
de Nucci
G
G
D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000032:analysis / Q000737:chemistry; D015951:Factor Xa / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D007915:Lepidoptera / Q000737:chemistry* / Q000254:growth & development*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data
D004220:Disulfides; D015951:Factor Xa