Identification of a functionally relevant signal peptide of mouse ficolin A.
Mouse ficolin A is a plasma protein with lectin activity, and plays a role in host defense by binding carbohydrates, especially GlcNAc, on microorganisms. The ficolin A subunit consists of an N-terminal signal peptide, a collagen-like domain, and a C-terminal fibrinogen-like domain. In this study, we show that ficolin A can be synthesized and oligomerized in a cell and secreted into culture medium. We also identify a functionally relevant signal peptide of ficolin A by using MS/MS analysis to determine the N-terminal sequence of secreted ficolin A. When the signal peptide of mouse ficolin A was fused with enhanced green fluorescent protein (EGFP), EGFP was released into HEK 293 cell medium, suggesting that the signal peptide can efficiently direct ficolin A secretion. Moreover, our results suggest that the signal peptide of ficolin A has potential application for the production of useful secretory proteins.
Department of Microbiology, College of Medicine, Hallym University, Chuncheon 200-702, Korea.
Sanghoon
S
Kwon
Min-Soo
MS
Kim
Dongbum
D
Kim
Keun-Wook
KW
Lee
Soo Young
SY
Choi
Jinseu
J
Park
Yeon Hyang
YH
Kim
Younghee
Y
Lee
Hyung-Joo
HJ
Kwon
D000595:Amino Acid Sequence; D000818:Animals; D002241:Carbohydrates; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D021382:Protein Sorting Signals; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D020539:Sequence Analysis, Protein; D000097604:Ficolins
D002241:Carbohydrates; D049452:Green Fluorescent Proteins; D037102:Lectins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000097604:Ficolins; C485184:enhanced green fluorescent protein