B38-CAP is a bacteria-derived ACE2-like enzyme that suppresses hypertension and cardiac dysfunction

Takafumi Minato,Midori Hoshizaki,Satoru Nirasawa,Yoshikazu Nakaoka,Tomokazu Yamaguchi,Souichi Koyota,Hiroyuki Watanabe,Saki Yokota,Taku Yoshiya,Miyuki Natsui,Yumiko Imai,Takeya Sato ,Tetsuo Gotoh ,Kumiko Yoshizawa‐Kumagaye ,Josef Penninger ,Takuya Inagaki ,Saori Takahashi,Tadashi Yoshihashi,Satoru Motoyama,Kazuhiko Nakahara,Ryo Ozawa,Keiji Kuba

doi:10.1038/s41467-020-14867-z

Abstract

Angiotensin-converting enzyme 2 (ACE2) is critically involved in cardiovascular physiology and pathology, and is currently clinically evaluated to treat acute lung failure. Here we show that the B38-CAP, a carboxypeptidase derived from Paenibacillus sp. B38, is an ACE2-like enzyme to decrease angiotensin II levels in mice. In protein 3D structure analysis, B38-CAP homolog shares structural similarity to mammalian ACE2 with low sequence identity. In vitro, recombinant B38-CAP protein catalyzed the conversion of angiotensin II to angiotensin 1–7, as well as other known ACE2 target peptides. Treatment with B38-CAP suppressed angiotensin II-induced hypertension, cardiac hypertrophy, and fibrosis in mice. Moreover, B38-CAP inhibited pressure overload-induced pathological hypertrophy, myocardial fibrosis, and cardiac dysfunction in mice. Our data identify the bacterial B38-CAP as an ACE2-like carboxypeptidase, indicating that evolution has shaped a bacterial carboxypeptidase to a human ACE2-like enzyme. Bacterial engineering could be utilized to design improved protein drugs for hypertension and heart failure.

Highlights

HPLC analyses for the metabolites of each peptide after B38-CAP treatment are shown in Supplementary Fig. 4a. *Cleavability of the peptides by ACE2 is from ref. 6
We examined whether B38CAP treatment affects Ang II-induced elevation of BP
It had been reported that an immune response is associated with the chronic infusion of rhACE2 resulting in the degradation of rhACE226, this was not observed for B38-CAP; there were no antibodies against B38-CAP detectable in the serum of mice infused with B38-CAP for 2 weeks (Supplementary Fig. 8c)

Summary

Methods

Sequence comparison was performed using BLAST and MEROPS (http://merops.sanger.ac.uk/) tools available online. Similarity search and superposition of a 3D structure of proteins was performed by Molecular Operating Environment (MOE 2016.08; Chemical Computing Group, Inc., Montreal, QC, Canada). Multiple sequence alignment between enzyme sequences was performed using the CLUSTALW tool available online. Phylogenetic tree drawing was performed using NJplot program Genomic DNAs were isolated from Paenibacillus sp. Subtilis NBRC 13719, and B. amyloliquefaciens NBRC 3022 (Supplementary Table 1). Expression plasmids, which encode B38-CAP, BS-CAP, b

Results

Discussion

Conclusion