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Titolo:
CHARACTERIZATION OF HUMAN LACTOFERRIN PRODUCED IN THE BACULOVIRUS EXPRESSION SYSTEM
Autore:
SALMON V; LEGRAND D; GEORGES B; SLOMIANNY MC; CODDEVILLE B; SPIK G;
Indirizzi:
UNIV SCI & TECH LILLE FLANDRES ARTOIS,CHIM BIOL LAB F-59655 VILLENEUVE DASCQ FRANCE UNIV SCI & TECH LILLE FLANDRES ARTOIS,CHIM BIOL LAB F-59655 VILLENEUVE DASCQ FRANCE UNIV SCI & TECH LILLE FLANDRES ARTOIS,UMR CNRS 111 F-59655 VILLENEUVEDASCQ FRANCE INST PASTEUR,LAB IMMUNOL CELLULAIRE,URA CNRS 1854 F-59019 LILLE FRANCE
Titolo Testata:
Protein expression and purification
fascicolo: 2, volume: 9, anno: 1997,
pagine: 203 - 210
SICI:
1046-5928(1997)9:2<203:COHLPI>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
RECOMBINANT HUMAN LACTOFERRIN; COLONY-STIMULATING ACTIVITY; GAS-LIQUID CHROMATOGRAPHY; IRON-BINDING PROTEINS; HUMAN LACTOTRANSFERRIN; NUCLEOTIDE-SEQUENCE; METHYL GLYCOSIDES; ESCHERICHIA-COLI; BRUSH-BORDER; CELL-LINES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
44
Recensione:
Indirizzi per estratti:
Citazione:
V. Salmon et al., "CHARACTERIZATION OF HUMAN LACTOFERRIN PRODUCED IN THE BACULOVIRUS EXPRESSION SYSTEM", Protein expression and purification, 9(2), 1997, pp. 203-210

Abstract

Lactoferrin, an iron-binding 80-kDa glycoprotein, is a major component of human milk whose structure is now well defined. The binding site of lactoferrin to the membrane receptor of lymphocyte has been locatedin the region 4-52, but the amino acids directly involved in the interaction have not been identified yet. To gain further insights into the structure-function relationships of the lactoferrin binding site, wefirst expressed the cDNA encoding human lactoferrin in the lepidoptera Spodoptera frugiperda cells (Sf9) using a recombinant baculovirus. The selected transformant secreted an N-glycosylated protein of 78 kDa which was immunoprecipitated by specific anti-lactoferrin antibodies. To confirm the structure and the function of the recombinant lactoferrin, the protein was purified by ion-exchange chromatography and its physical, biochemical, and biological properties were compared with those of the native protein. In particular, the N-terminal amino acid sequence and the iron-binding stability as a function of pH, of both proteins, were identical. The main difference concerns the glycosylation which leads to glycans of lower molecular masses as detected by the electrophoretic mobility of lactoferrin after N-glycosidase F treatment and matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. Despite the different glycosylation features, the recombinant lactoferrin retained the binding property to the Jurkat human lymphoblastic T-cell line of the native lactoferrin. On the basis of these analyses, production of protein mutants generated by site-directed mutagenesis is now in process. (C) 1997 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 12/07/20 alle ore 06:30:23