Structures and function of the amino acid polymerase cyanophycin synthetase

Sharon, Itai; Haque, Asfarul S.; Grogg, Marcel; Lahiri, Indrajit; Seebàch, Dieter; Leschziner, Andres E.; Hilvert, Donald; Schmeing, T.M.

doi:10.1038/s41589-021-00854-y

Cited by 29 publications

(104 citation statements)

References 58 publications

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“…Therefore, M core assumes a distinct catalytic site from the G domain in Te CphA1, which is similar to the structural observation of Su CphA1 13 . On the other hand, the structural evidence of the ATP-bound forms of the G and M domains remains insufficient for the CphA1 enzymes from Acinetobacter baylyi DSM587 ( Ab CphA1) and Tatumella morbirosei DSM23827 ( Tm CphA1) 13 . M lid is not well resolved in the apo- and ATPγS-bound states (Fig.…”

Section: Resultssupporting

confidence: 82%

“…The cryo-EM data of the substrate-bound state were collected under a high concentration of l -aspartate (100 mM), cyanophycin primer peptide, (β-Asp-Arg) 4 , and ATPγS. Te CphA1 consists of three domains: an N-terminal domain (N domain, residues 1‒161) that adopts a partially similar structure to E. coli RNA-polymerase α-subunit 13 , a glutathione synthetase-like domain (G domain, residues 162‒487), and a MurE-like muramyl ligase domain (M domain, residues 488‒876) (Fig. 1b ).…”

Section: Resultsmentioning

confidence: 99%

“…CphA1 uses a low-molecular-weight cyanophycin (at least 3‒4 dipeptides long) as a primer for cyanophycin polymerization 12 . Recently, the tertiary structure of CphA1 has become available and reveals that amino acid polymerization is driven by three distinct domains 13 : the N-terminal domain (N domain), the middle glutathione synthetase-like domain (G domain) with the ATP-grasp fold, and the MurE-like muramyl ligase domain (M domain). The discrete active sites of the G and M domains are used for extending the Asp backbone and attaching the Arg sidechain, respectively (Supplementary Fig.…”

Section: Introductionmentioning

confidence: 99%

“…1 ). The N domain has been proposed to anchor cyanophycin polymers through electrostatic interactions 13 . However, it remains unclear how ATP-dependent addition of aspartate to cyanophycin is initiated at the catalytic site of the G domain.…”

Section: Introductionmentioning

confidence: 99%

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Structural bases for aspartate recognition and polymerization efficiency of cyanobacterial cyanophycin synthetase

et al. 2022

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Cyanophycin is a natural biopolymer consisting of equimolar amounts of aspartate and arginine as the backbone and branched sidechain, respectively. It is produced by a single enzyme, cyanophycin synthetase (CphA1), and accumulates as a nitrogen reservoir during N2 fixation by most cyanobacteria. A recent structural study showed that three constituent domains of CphA1 function as two distinct catalytic sites and an oligomerization interface in cyanophycin synthesis. However, it remains unclear how the ATP-dependent addition of aspartate to cyanophycin is initiated at the catalytic site of the glutathione synthetase-like domain. Here, we report the cryogenic electron microscopy structures of CphA1, including a complex with aspartate, cyanophycin primer peptide, and ATP analog. These structures reveal the aspartate binding mode and phosphate-binding loop movement to the active site required for the reaction. Furthermore, structural and mutational data show a potential role of protein dynamics in the catalytic efficiency of the arginine condensation reaction.

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Section: Resultssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural bases for aspartate recognition and polymerization efficiency of cyanobacterial cyanophycin synthetase

et al. 2022

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“…Cyanophycin is synthesized by the enzyme cyanophycin synthetase (CphA), which requires a primer, Mg 2+ , K + , ATP and two amino acids for its catalytic action ( Simon, 1976 ). CphA uses a short peptide as a primer and then extends it to produce cyanophycin ( Berg et al, 2000 ; Sharon et al, 2021 ). The α-carboxylic group of aspartic acid in the backbone is activated by phosphorylation, whereby ATP is converted to ADP.…”

Section: Introductionmentioning

confidence: 99%

Cyanophycin Modifications—Widening the Application Potential

Kwiatos

Steinbüchel

2021

Front. Bioeng. Biotechnol.

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A circular bioeconomy approach is essential to slowing down the fearsome ongoing climate change. Replacing polymers derived from fossil fuels with biodegradable biobased polymers is one crucial part of this strategy. Cyanophycin is a polymer consisting of amino acids produced by cyanobacteria with many potential applications. It consists mainly of aspartic acid and arginine, however, its composition may be changed at the production stage depending on the conditions of the polymerization reaction, as well as the characteristics of the enzyme cyanophycin synthetase, which is the key enzyme of catalysis. Cyanophycin synthetases from many sources were expressed heterologously in bacteria, yeast and plants aiming at high yields of the polymer or at introducing different amino acids into the structure. Furthermore, cyanophycin can be modified at the post-production level by chemical and enzymatic methods. In addition, cyanophycin can be combined with other compounds to yield hybrid materials. Although cyanophycin is an attractive polymer for industry, its usage as a sole material remains so far limited. Finding new variants of cyanophycin may bring this polymer closer to real-world applications. This short review summarizes all modifications of cyanophycin and its variants that have been reported within the literature until now, additionally addressing their potential applications.

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Recombinant Multi‐l‐Arginyl‐Poly‐l‐Aspartate (Cyanophycin) as an Emerging Biomaterial

Tseng

Fang

2023

Macromolecular Bioscience

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Multi‐l‐arginyl‐poly‐l‐aspartate (MAPA) is a non‐ribosomal polypeptide which synthesis is directed by cyanophycin synthetase, and its production can be achieved using recombinant microorganisms carrying the cphA gene. Along its poly‐aspartate backbone, arginine or lysine links to each aspartate via an isopeptide bond. MAPA is a zwitterionic polyelectrolyte full of charged carboxylic, amine, and guanidino groups. In aqueous solution, MAPA exhibits dual thermal and pH responses similar to those stimuli‐responsive polymers. Being biocompatible, the films containing MAPA can support cell proliferation and elicits minimal immune response in macrophages. Dipeptides from MAPA after enzymatic treatments can provide nutritional benefits. In light of the increasing interest in MAPA, this article focuses on the recent discovery of the function of cyanophycin synthetase and the potentials of MAPA as a biomaterial.

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Structures and function of the amino acid polymerase cyanophycin synthetase

Cited by 29 publications

References 58 publications

Structural bases for aspartate recognition and polymerization efficiency of cyanobacterial cyanophycin synthetase

Structural bases for aspartate recognition and polymerization efficiency of cyanobacterial cyanophycin synthetase

Cyanophycin Modifications—Widening the Application Potential

Recombinant Multi‐l‐Arginyl‐Poly‐l‐Aspartate (Cyanophycin) as an Emerging Biomaterial

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