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Bioscience Reports (2010) 30, (1–10) (Printed in Great Britain)

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Original Paper

C12ORF39, a novel secreted protein with a typical amidation processing signal

Bingbing Wan*†‡, Xin‑Rong Wang‡, Yu‑Bo Zhou‡, Xin Zhang‡, Keke Huo*†¹ and Ze‑Guang Han*†‡¹

*State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China, †Chinese National Human Genome Center at Shanghai, Shanghai 201203, China, and ‡Shanghai-Ministry Key Laboratory of Disease and Health Genomics, Chinese National Human Genome Center at Shanghai, Shanghai 201203, China

Key words: amidation/proteolytic processing signal, antibody production, chromosome 12 open-reading framework 39 (C12ORF39), placenta, post-translational modification, secreted protein.

Abbreviations used: BFA, brefeldin A; C12ORF39, chromosome 12 open-reading framework 39; DMEM, Dulbecco's modified Eagle's medium; ER, endoplasmic reticulum; hGH, human growth hormone; GST, glutathione transferase; Ni–NTA, Ni²⁺–nitrilotriacetate; RT-PCR, reverse transcription-PCR; TRX, thioredoxin; YFP, yellow fluorescence protein.

¹Correspondence may be addressed to either of these authors (email kkhuo@fudan.edu.cn or hanzg@chgc.sh.cn).

Synopsis

In the present study we describe a novel secreted protein, named C12ORF39 (chromosome 12 open-reading framework 39), which contains a typical amidation/proteolytic processing signal (Gly–Arg–Arg motif). Interestingly, C12ORF39 protein is not hydrolysed, but is a full-length protein without signal peptides. Western blotting indicated that c-Myc-tagged C12ORF39 is secreted into culture medium in transfected HeLa cells. Quantitative RT-PCR (reverse transcription-PCR) analysis revealed that c12orf39 is mainly expressed in placenta and brain. Immunohistochemistry on formalin-fixed paraffin-embedded human term placenta using a rabbit antibody against human C12ORF39 demonstrated that the protein was localized extracellularly, surrounding the trophoblastic cells. In addition, C12ORF39 secretion could be blocked by brefeldin A, suggesting that the secretion of C12ORF39 is dependent on the Golgi apparatus. Furthermore, laser-scanning confocal microscopy also confirmed that the C12ORF39 protein co-localized with the Golgi apparatus. Taken together, although C12ORF39 is not a secreted small peptide, it can also be secreted to play a role in the biological functions of the placenta.

Received 17 October 2008/12 January 2009; accepted 4 February 2009

Published as Immediate Publication 4 February 2009, doi:10.1042/BSR20080156

© The Authors Journal compilation © 2010 Biochemical Society