A cryptic third active site in cyanophycin synthetase creates primers for polymerization

Sharon, Itai; Pinus, Sharon; Grogg, Marcel; Moitessier, Nicolas; Hilvert, Donald; Schmeing, T. Martin

doi:10.1038/s41467-022-31542-7

A cryptic third active site in cyanophycin synthetase creates primers for polymerization

Journal Article · Thu Jul 07 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-31542-7· OSTI ID:1904103

^[1]; Pinus, Sharon ^[2]; Grogg, Marcel ^[3]; ^[2]; ^[3]; ^[2]

McGill University, Montreal, QC (Canada); OSTI
McGill University, Montreal, QC (Canada)
Eidgenoessische Technische Hochschule (ETH), Zurich (Switzerland)

Cyanophycin is a nitrogen reserve biopolymer in many bacteria that has promising industrial applications. Made by cyanophycin synthetase 1 (CphA1), it has a poly-L-Asp backbone with L-Arg residues attached to each aspartate sidechain. CphA1s are thought to typically require existing segments of cyanophycin to act as primers for cyanophycin polymerization. In this study, we show that most CphA1s will not require exogenous primers and discover the surprising cause of primer independence: CphA1 can make minute quantities of cyanophycin without primer, and an unexpected, cryptic metallopeptidase-like active site in the N-terminal domain of many CphA1s digests these into primers, solving the problem of primer availability. We present co-complex cryo-EM structures, make mutations that transition CphA1s between primer dependence and independence, and demonstrate that primer dependence can be a limiting factor for cyanophycin production in heterologous hosts. In CphA1, domains with opposite catalytic activities combine into a remarkable, self-sufficient, biosynthetic nanomachine.

View Accepted Manuscript (DOE)

Research Organization:: USDOE Joint Genome Institute (JGI), Berkeley, CA (United States)

Sponsoring Organization:: Canadian Institutes of Health Research; ETH Zurich; Schweizerischer Nationalfonds; USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1904103

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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