Comparison of different signal peptides for protein secretion in nonlytic insect cell system.
Protein expression and secretion in insect cells have been widely studied in the baculovirus-infected insect cell system. In directly transfected insect cells only intracellular expression and purification of recombinant proteins have been studied in detail. To examine multiple recombinant protein variants, easy and fast expression and a purification screening system are required. The aim of this study was to establish an effective and rapid secretion system for human azurocidin using directly transfected insect cells. We also constructed and tested expression vectors possessing heterologous signal peptides derived from human azurocidin, yellow lupin diphosphonucleotide phosphatase/phosphodiesterase (PPD1), and papaya papain IV to secrete yellow lupin and red kidney bean purple acid phosphatases, PPD1, and papain IV. Our results demonstrate that the secretion vectors used here can direct recombinant proteins to the culture medium very effectively, allowing their simple purification on a small/medium scale. Based on secretion and activity analyses it seems that the azurocidin signal peptide is one of the most potent secretion signals.
Olczak
Mariusz
M
Laboratory of Biochemistry, Institute of Biochemistry and Molecular Biology, Wroclaw University, 50-137 Wroclaw, Poland. Mariusz.Olczak@bf.uni.wroc.pl
Olczak
Teresa
T
D000135:Acid Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001798:Blood Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002478:Cells, Cultured / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D003470:Culture Media; D005822:Genetic Vectors / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007313:Insecta / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000096:biosynthesis / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000502:physiology; D021382:Protein Sorting Signals / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology; D014162:Transfection / Q000379:methods
C062991:AZU1 protein, human; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D002352:Carrier Proteins; D003470:Culture Media; D006023:Glycoproteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C091137:purple acid phosphatase; D000135:Acid Phosphatase; D010727:Phosphoric Diester Hydrolases