Characterization of partially truncated human midkine expressed in Pichia pastoris.
Recombinant human midkine (rh-midkine) was expressed under the control of the AOX1 gene promoter in Pichiapastoris. Approximately 640 mg of rh-midkine was secreted into one liter of medium of the high cell-density fermentation. The protein processing of the rh-midkine was done efficiently and correctly in P. pastoris, and O-mannosylation was not detected in the purified rh-midkine. However, only about 30% of the purified rh-midkine was intact. The other ones lost 5-12 amino acid residues from the amino-termini, provably by proteolysis. Even the mixture of these truncated midkines could promote CHO cell proliferation as well as the authentic rh-midkine.
Murasugi
Akira
A
Meiji Cell Technology Center, Meiji Milk Products Co, Ltd, Odawara, Kanagawa, Japan. AKIRA_MURASUGI@MEIJI-MILK.COM
Tohma-Aiba
Yumiko
Y
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D006224:Cricetinae; D016207:Cytokines; D005285:Fermentation; D015870:Gene Expression; D006801:Humans; D013058:Mass Spectrometry; D000078792:Midkine; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D013997:Time Factors
D002352:Carrier Proteins; D016207:Cytokines; D011994:Recombinant Proteins; D000078792:Midkine