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Open AccessArticle

Purification and Characterization of a New CRISP-Related Protein from Scapharca broughtonii and Its Immunomodulatory Activity

1
Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China
2
Department of Pharmacology, College of Pharmacy, Jinan University, Guangzhou 510632, China
3
Department of Natural Product Chemistry, College of Pharmacy, Jinan University, Guangzhou 510632, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Mar. Drugs 2020, 18(6), 299; https://doi.org/10.3390/md18060299
Received: 17 May 2020 / Revised: 31 May 2020 / Accepted: 2 June 2020 / Published: 4 June 2020
More and more attention has been paid to bioactive compounds isolated from marine organisms or microorganisms in recent years. At the present study, a new protein coded as HPCG2, was purified from Scapharca broughtonii by stepwise chromatography methods. The molecular weight of HPCG2 was determined to be 30.71 kDa by MALDI-TOF-MS. The complete amino acid sequence of HPCG2 was obtained by tandem mass spectrometry combined with transcriptome database analysis, and its secondary structure was analyzed using circular dichroism. HPCG2 comprised 251 amino acids and contained 28.4% α-helix, 26% β-sheet, 18.6% β-turn, and 29.9% random coil. HPCG2 was predicted to be a cysteine-rich secretory protein-related (CRISP-related) protein by domain prediction. Moreover, HPCG2 was proved to possess the immunomodulatory effect on the murine immune cells. MTT assay showed that HPCG2 promoted the proliferation of splenic lymphocytes and the cytotoxicity of NK cells against YAC-1 cells. Flow cytometry test revealed that HPCG2 enhanced the phagocytic function of macrophages and polarized them into M1 type in RAW264.7 cells. In particular, Western blot analysis indicated that the immunomodulatory mechanism of HPCG2 was associated with the regulation on TLR4/JNK/ERK and STAT3 signaling pathways in RAW 264.7 cells. These results suggested that HPCG2 might be developed as a potential immunomodulatory agent or new functional product from marine organisms. View Full-Text
Keywords: Scapharca broughtonii protein; purification; structural characterization; CRISP-related protein; immunomodulatory activity Scapharca broughtonii protein; purification; structural characterization; CRISP-related protein; immunomodulatory activity
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MDPI and ACS Style

Liu, W.; Bi, S.; Li, C.; Zheng, H.; Guo, Z.; Luo, Y.; Ou, X.; Song, L.; Zhu, J.; Yu, R. Purification and Characterization of a New CRISP-Related Protein from Scapharca broughtonii and Its Immunomodulatory Activity. Mar. Drugs 2020, 18, 299.

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