TY - JOUR
T1 - Cloning and recombinant expression of a barred sand bass (Paralabrax nebulifer) cDNA. The encoded protein displays structural homology and immunological crossreactivity to human complement/cofactor related plasma proteins
AU - Zipfel, Peter F.
AU - Kemper, Claudia
AU - Dahmen, Antje
AU - Gigli, Irma
N1 - Funding Information:
qA bstracL-A new cofactor related cDNA in the bony fish Paralablax nebulifr, (barred sand bass) was isolated from a sand bass liver cDNA library. The clone (~71) is 1040 bp in size and the predicted translation product of 204 amino acids contains a hydrophobic signal peptide, which is followed by a region of three short consensus repeats (SCRs). The three SCRs display high homology to SCRs of the 110 kDa chain of the sand bass plasma cofactor protein, and to a lesser degree to human complement factor H related protein 3 (FHR-3) and to human factor H. Recombinant expression of the c71 cDNA in the baculovirus system shows a product of an apparent molecular mass of 27 kDa, which is secreted and glycosylated. It also contains a His-tag *This work was supported by NIH grant AI20067 and a grant from the Deutsche Forschungsgemeinschaft (Zi432/2).The nucleotide sequence reported in this paper has been submitted to the Gen Bank@/EMBL Data Bank with accession number.This work is part of the doctoral thesis of C.K. at the Depatment of Biology at the University of Hamburg. Address correspondence to Dr P. F. Zipfel, Bernhard Nocht Institute, Bernhard Nocht Strasse 74, 20359 Hamburg, Germany.
PY - 1996/11
Y1 - 1996/11
N2 - A new cofactor related cDNA in the bony fish Paralablax nebulifer, (barred sand bass) was isolated from a sand bass liver cDNA library. The clone (c71) is 1040 bp in size and the predicted translation product of 204 amino acids contains a hydrophobic signal peptide, which is followed by a region of three short consensus repeats (SCRs). The three SCRs display high homology to SCRs of the 110 kDa chain of the sand bass plasma cofactor protein, and to a lesser degree to human complement factor H related protein 3 (FHR-3) and to human factor H. Recombinant expression of the c71 cDNA in the baculovirus system shows a product of an apparent molecular mass of 27 kDa, which is secreted and glycosylated. It also contains a His-tag for purification purposes. Removal of the His-tag yields a 24 kDa protein, and deglycosylation further reduces the molecular mass to 21 kDa. This size is in agreement with the calculated molecular mass based on amino acid composition. The sand bass SBCFR-1 protein is immunologically related to the human complement proteins, factor H and factor H-related protein 3. The recombinantly expressed protein reacted with antisera against the human FHR- 3 protein and SCRs 19-20 of human factor H. The presence of SCR-containing proteins in sand bass plasma and their structural and immunological homology to human FHR-3 and factor H suggests for n common function between these evolutionary related proteins.
AB - A new cofactor related cDNA in the bony fish Paralablax nebulifer, (barred sand bass) was isolated from a sand bass liver cDNA library. The clone (c71) is 1040 bp in size and the predicted translation product of 204 amino acids contains a hydrophobic signal peptide, which is followed by a region of three short consensus repeats (SCRs). The three SCRs display high homology to SCRs of the 110 kDa chain of the sand bass plasma cofactor protein, and to a lesser degree to human complement factor H related protein 3 (FHR-3) and to human factor H. Recombinant expression of the c71 cDNA in the baculovirus system shows a product of an apparent molecular mass of 27 kDa, which is secreted and glycosylated. It also contains a His-tag for purification purposes. Removal of the His-tag yields a 24 kDa protein, and deglycosylation further reduces the molecular mass to 21 kDa. This size is in agreement with the calculated molecular mass based on amino acid composition. The sand bass SBCFR-1 protein is immunologically related to the human complement proteins, factor H and factor H-related protein 3. The recombinantly expressed protein reacted with antisera against the human FHR- 3 protein and SCRs 19-20 of human factor H. The presence of SCR-containing proteins in sand bass plasma and their structural and immunological homology to human FHR-3 and factor H suggests for n common function between these evolutionary related proteins.
UR - http://www.scopus.com/inward/record.url?scp=0030292831&partnerID=8YFLogxK
U2 - 10.1016/S0145-305X(96)00025-0
DO - 10.1016/S0145-305X(96)00025-0
M3 - Journal articles
C2 - 9040983
AN - SCOPUS:0030292831
SN - 0145-305X
VL - 20
SP - 407
EP - 416
JO - Developmental and Comparative Immunology
JF - Developmental and Comparative Immunology
IS - 6
ER -
