Recombinant porcine prorelaxin produced in Chinese hamster ovary cells is biologically active.
Although prorelaxin has a similar structure as proinsulin, the posttranslational processing of prorelaxin seems to be quite different from that of proinsulin. There are no pairs of basic residues flanking the relaxin moiety in most prorelaxins studied so far. Instead, the prorelaxins of many species contains a tetrabasic sequence (Arg-Lys-Lys-Arg) between the connecting peptide and the A-chain. This is the recognition sequence of furin. In order to study this possible processing by furin, we express the recombinant porcine prorelaxin in Chinese hamster ovary cells. The expected 19 kDa recombinant porcine prorelaxin was found to be constitutively secreted into the medium at a level of approximately 250 ng/ml. No conversion of the 19 kDa prorelaxin into the 6 kDa relaxin was observed. Unlike most prohormones which are biologically inactive, the recombinant prorelaxin was found to be biologically active in an in vitro bioassay.
Vu
A L
AL
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66160.
Green
C B
CB
Roby
K F
KF
Soares
M J
MJ
Fei
D T
DT
Chen
A B
AB
Kwok
S C
SC
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D006224:Cricetinae; D004717:Endometrium / Q000187:drug effects / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D012065:Relaxin / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D015183:Restriction Mapping; D013552:Swine; D014162:Transfection
D046911:Macromolecular Substances; D011498:Protein Precursors; D011994:Recombinant Proteins; C038656:prorelaxin; D012065:Relaxin