Identification and characterization of Scp15, a protein from Streptomyces coelicolor A3(2) inducing neurites in PC12 cells.
We previously showed that a fungal protein, p15, induces neurite outgrowth and differentiation of rat pheochromocytoma PC12 cells. We report here the identification and characterization of a protein similar to p15, found in Streptomyces coelicolor A3(2). This hypothetical protein, tentatively named Scp15, has significant similarity with p15, including conserved positions of four cysteine residues involved in the formation of essential disulfide bonds in p15. Hexahistidine-tagged recombinant Scp15 proteins were produced in Escherichia coli, purified, and analyzed for their neurite-inducing activity. Although they were less active than p15, they dose-dependently induced neurites and the expression of neurofilament M. Neurite outgrowth by Scp15 was inhibited by nicardipine, suggesting that Scp15 induces neurites via activation of a calcium signaling pathway.
Nakashima
Satoru
S
Department of Biotechnology, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Wakatsuki
Shuji
S
Yokoyama
Tatsuya
T
Arioka
Manabu
M
Kitamoto
Katsuhiko
K
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D015153:Blotting, Western; D002121:Calcium Channel Blockers / Q000494:pharmacology; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D016501:Neurites / Q000737:chemistry*; D016900:Neurofilament Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D009529:Nicardipine / Q000494:pharmacology; D016716:PC12 Cells; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013302:Streptomyces / Q000737:chemistry*
D001426:Bacterial Proteins; D002121:Calcium Channel Blockers; D004269:DNA, Bacterial; D016900:Neurofilament Proteins; D011994:Recombinant Proteins; C476805:Scp15 protein, Streptomyces coelicolor; D009529:Nicardipine