Identification of sites in apolipoprotein A-I susceptible to chymase and carboxypeptidase A digestion

• Published in: Bioscience reports 2012-12, Vol.33 (1), p.49-56
• Main Authors: Usami, Yoko, Kobayashi, Yukihiro, Kameda, Takahiro, Miyazaki, Akari, Matsuda, Kazuyuki, Sugano, Mitsutoshi, Kawasaki, Kenji, Kurihara, Yuriko, Kasama, Takeshi, Tozuka, Minoru
• Format: Article
• Language: English
• Subjects: Aorta, Thoracic - chemistry; Aorta, Thoracic - pathology; Apolipoprotein A-I - blood; Apolipoprotein A-I - chemistry; Biomarkers - blood; Carboxypeptidases A - chemistry; Catalytic Domain; Chymases - chemistry; Electrophoresis, Polyacrylamide Gel; Humans; Immunohistochemistry; Lipoproteins, HDL3 - blood; Lipoproteins, HDL3 - chemistry; Original Paper; Phenylalanine - chemistry; Plaque, Atherosclerotic - chemistry; Plaque, Atherosclerotic - pathology; Proteolysis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tyrosine - chemistry
• ISSN: 0144-8463; 1573-4935
• DOI: 10.1042/BSR20120094

MCs (mast cells) adversely affect atherosclerosis by promoting the progression of lesions and plaque destabilization. MC chymase cleaves apoA-I (apolipoprotein A-I), the main protein component of HDL (high-density lipoprotein). We previously showed that C-terminally truncated apoA-I (cleaved at the carboxyl side of Phe(225)) is present in normal human serum using a newly developed specific mAb (monoclonal antibody). In the present study, we aimed to identify chymase-induced cleavage sites in both lipid-free and lipid-bound (HDL(3)) forms of apoA-I. Lipid-free apoA-I was preferentially digested by chymase, at the C-terminus rather than the N-terminus. Phe(229) and Tyr(192) residues were the main cleavage sites. Interestingly, the Phe(225) residue was a minor cleavage site. In contrast, the same concentration of chymase failed to digest apoA-I in HDL(3); however, a 100-fold higher concentration of chymase modestly digested apoA-I in HDL(3) at only the N-terminus, especially at Phe(33). CPA (carboxypeptidase A) is another MC protease, co-localized with chymase in severe atherosclerotic lesions. CPA, in vitro, further cleaved C-terminal Phe(225) and Phe(229) residues newly exposed by chymase, but did not cleave Tyr(192). These results indicate that several forms of C-terminally and N-terminally truncated apoA-I could exist in the circulation. They may be useful as new biomarkers to assess the risk of CVD (cardiovascular disease).