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Titolo:
INTERACTION OF HIGH-MOLECULAR-WEIGHT BASIC FIBROBLAST GROWTH-FACTOR WITH ENDOTHELIUM - BIOLOGICAL-ACTIVITY AND INTRACELLULAR FATE OF HUMAN RECOMBINANT M(R) 24,000 BFGF
Autore:
GUALANDRIS A; URBINATI C; RUSNATI M; ZICHE M; PRESTA M;
Indirizzi:
UNIV BRESCIA,SCH MED,DEPT BIOMED SCI & BIOTECHNOL,GEN PATHOL & IMMUNOL UNIT BRESCIA ITALY UNIV BRESCIA,SCH MED,DEPT BIOMED SCI & BIOTECHNOL,GEN PATHOL & IMMUNOL UNIT BRESCIA ITALY UNIV FLORENCE,DEPT PHARMACOL FLORENCE ITALY
Titolo Testata:
Journal of cellular physiology
fascicolo: 1, volume: 161, anno: 1994,
pagine: 149 - 159
SICI:
0021-9541(1994)161:1<149:IOHBFG>2.0.ZU;2-#
Fonte:
ISI
Lingua:
ENG
Soggetto:
PLASMINOGEN-ACTIVATOR PRODUCTION; NIH 3T3 CELLS; DNA-SYNTHESIS; NH2-TERMINAL EXTENSION; NUCLEAR-LOCALIZATION; SIGNAL SEQUENCE; RECEPTOR; FORMS; MIGRATION; BINDING;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
49
Recensione:
Indirizzi per estratti:
Citazione:
A. Gualandris et al., "INTERACTION OF HIGH-MOLECULAR-WEIGHT BASIC FIBROBLAST GROWTH-FACTOR WITH ENDOTHELIUM - BIOLOGICAL-ACTIVITY AND INTRACELLULAR FATE OF HUMAN RECOMBINANT M(R) 24,000 BFGF", Journal of cellular physiology, 161(1), 1994, pp. 149-159

Abstract

The single-copy gene of human basic fibroblast growth factor (bFGF) encodes four co-expressed isoforms, with an apparent molecular weight (M(r)) of 24 kD, 22.5 kD, 22 kD, and 18 kD, co-translated from a singlemRNA. As a tool for the study of the role exerted by the different bFGF isoforms in the biology of endothelial cells, human recombinant 24-kD bFGF was produced and purified from transformed Escherichia coli cells. To this purpose, the novel CUG start codon present in human bFGF cDNA and responsible for the synthesis of 24-kD bFGF was mutagenized to the classic AUG start codon. Transient expression of the mutagenizedcDNA in simian COS-1 cells, followed by immunolocalization and subcellular fractionation, resulted in the synthesis of high levels of 24-kDbFGF, which localizes in the cell nucleus as an intact protein. When the same 24-kD bFGF cDNA was expressed in E. coli, the recombinant protein was purified to homoge neity by heparin-Sepharose and ion-exchange chromatography. Recombinant 24-kD bFGF was similar to recombinant 18-kD bFGF in receptor-binding activity and in inducing cell proliferation, plasminogen activator production, and chemotactic movement in cultured endothelial cells. In agreement with the in vitro observations, 24-kD bFGF and 18-kD bFGF exerted a similar angiogenic response when assayed in vivo in the rabbit cornea. Experiments performed with the radiolabeled molecule demonstrated that 24-kD bFGF has an intrinsic ability to bind to high-affinity receptors when added to endothelial GM 7373 cell cultures. Receptor-bound 24-kD bFGF is internalized within the cell and associates with the nucleus with kinetics similar to 18-kD bFGF. Internalized 24-kD bFGF is first processed to the 18-kD form via achloroquine-insensitive pathway and then to smaller fragments into the lysosomal compartment. At variance with the data obtained in transfected COS-1 cells, only limited amounts of exogenous internalized 24-kDbFGF associates with the nucleus in the intact form, mostly of the nuclear-bound molecule being represented by the processed 18-kD protein and by smaller degradation products. In conclusion, human recombinant 24-kD bFGF exerts a biological response in endothelial cells similar to 18-kD bFGF both in vitro and in vivo. Our data point to a different intracellular behavior of the high-molecular-weight bFGF isoform when added exogenously to cultured cells or when produced endogenously in transfected cells. (C) 1994 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 00:59:02