Expression and characterization of a B cell growth promoting polypeptide derived from the 12 kDa B cell growth factor gene (BCGF 1).
The expression and partial purification of recombinant 12 kDa B cell growth factor are reported. The polypeptide was derived from the genomic sequence of the gene (BCGF 1) which is here shown to be a single copy gene that localizes to human chromosome 16. When expressed as a glutathione S-transferase fusion protein in E. coli, the protein appears as a 38 kDa polypeptide in Western blot analysis using a peptide antibody. The purified fusion protein stimulates the proliferation of activated human B cells in a dose-dependent manner, and the active site resides within the 104 carboxy-terminal amino acids. The availability of biologically active recombinant 12 kDa B cell growth factor will enable its evaluation in B cell growth regulation, and provides a new means of in vitro culturing of human B lymphocytes.
Kovanen
P E
PE
Department of Pathology, University of Helsinki, Finland.
Virtaneva
K
K
Harju
L
L
Kantor
C
C
Gahmberg
C G
CG
Timonen
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D001483:Base Sequence; D001665:Binding Sites; D015139:Blotting, Southern; D015153:Blotting, Western; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002885:Chromosomes, Human, Pair 16; D004247:DNA / Q000097:blood / Q000302:isolation & purification; D017931:DNA Primers; D015870:Gene Expression; D005982:Glutathione Transferase / Q000096:biosynthesis; D006801:Humans; D006822:Hybrid Cells; D015847:Interleukin-4 / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D007962:Leukocytes / Q000378:metabolism*; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D015183:Restriction Mapping; D012377:Rodentia; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D015847:Interleukin-4; D004247:DNA; D005982:Glutathione Transferase