Introduction of a low molecular weight agonist peptide for complement C3a receptor into soybean proglycinin A1aB1b subunit by site-directed mutagenesis.
LPLPR, a complement C3a agonist peptide, with hypocholesterolemic activity was introduced into the homologous site of soybean proglycinin A1aB1b subunit by site-directed mutagenesis. This modified proglycinin was expressed in E. coli and recovered from the insoluble fraction. LPLPR was released by the action of chymotrypsin and trypsin as expected. Furthermore, two peptides (RPSYLPLPR and PSYLPLPR) with extended sequence at the amino-terminus of LPLPR were obtained. Their ileum-contracting activity was 9 to approximately 13 times stronger than that of LPLPR. The overall yields of purified LPLPR, RPSYLPLPR and PSYLPLPR were 25%, 12%, and 0.7% respectively.
Takenaka
Y
Y
Division of Food Bioscience, Graduate School of Agriculture, Kyoto University, Uji, Japan.
Utsumi
S
S
Yoshikawa
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D006168:Guinea Pigs; D007082:Ileum / Q000187:drug effects / Q000502:physiology; D066298:In Vitro Techniques; D008565:Membrane Proteins; D008970:Molecular Weight; D009119:Muscle Contraction / Q000187:drug effects; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D011951:Receptors, Complement / Q000819:agonists*; D017386:Sequence Homology, Amino Acid; D030262:Soybean Proteins; D013025:Glycine max
D017931:DNA Primers; D008565:Membrane Proteins; D009842:Oligopeptides; D010940:Plant Proteins; D011951:Receptors, Complement; D030262:Soybean Proteins; C085567:complement C3a receptor; C438909:leucyl-prolyl-leucyl-prolyl-arginine; C405345:proglycinin A(1a)B(1b), soybean