Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 May 4;22(5):3271-3276.
doi: 10.1021/acs.cgd.2c00108. Epub 2022 Feb 22.

Protein Frameworks with Thiacalixarene and Zinc

Ronan J Flood  1 , Kiefer O Ramberg  1 , Darius B Mengel  1 , Francesca Guagnini  1 , Peter B Crowley  1
Affiliations

Protein Frameworks with Thiacalixarene and Zinc

Ronan J Flood et al. Cryst Growth Des. .

Abstract

Controlled protein assembly provides a means to generate biomaterials. Synthetic macrocycles such as the water-soluble sulfonato-calix[n]arenes are useful mediators of protein assembly. Sulfonato-thiacalix[4]arene (tsclx 4 ), with its metal-binding capacity, affords the potential for simultaneous macrocycle- and metal-mediated protein assembly. Here, we describe the tsclx 4 -/Zn-directed assembly of two proteins: cationic α-helical cytochrome c (cyt c) and neutral β-propeller Ralstonia solanacearum lectin (RSL). Two co-crystal forms were obtained with cyt c, each involving multinuclear zinc sites supported by the cone conformation of tsclx 4 . The tsclx 4 /Zn cluster acted as an assembly node via both lysine encapsulation and metal-mediated protein-protein contacts. In the case of RSL, tsclx 4 adopted the 1,2-alternate conformation and supported a dinuclear zinc site with concomitant encapsulation and metal-binding of two histidine side chains. These results, together with the knowledge of thiacalixarene/metal nanoclusters, suggest promising applications for thiacalixarenes in biomaterials and MOF fabrication.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Schematic structures of sclx4 and tsclx4.
Figure 2
Figure 2
Representative co-crystals of cyt c and tsclx4 in the presence of magnesium or zinc. Scale bar is 100 μm. See Figure S1 for alternative zinc-containing crystals.
Figure 3
Figure 3
Detail of the tsclx4/Zn complexes bound to cyt c in (A) Form I and (B) Form II. In each case, three protein chains (pink, green, and gray) are involved. Thiacalixarene, some key residues, and phosphates are shown as sticks. Note that His39 in Form I is omitted for clarity. The lower panels show the corresponding crystal packing, viewed along the b and c cell axes (drawn to scale) with proteins represented as gray ribbons and tsclx4/Zn/phosphate clusters as colored spheres.
Figure 4
Figure 4
(A) RSL-N23H forms a tetrahedral assembly mediated by His-Zn-His interactions and one tsclx4-/Zn-mediated interface. RSL trimers are gray, green, purple, and beige. (B) Detail of the tsclx4—dinuclear zinc species, with the 1,2-alternate conformation complexing two His side chains that coordinate two zinc ions.
See this image and copyright information in PMC

References

    1. Zhu J.; Avakyan N.; Kakkis A.; Hoffnagle A. M.; Han K.; Li Y.; Zhang Z.; Choi T. S.; Na Y.; Yu C.-J.; Tezcan F. A. Protein assembly by design. Chem. Rev. 2021, 121, 13701–13796. 10.1021/acs.chemrev.1c00308. - DOI - PMC - PubMed
    1. Kojima M.; Abe S.; Ueno T. Engineering of protein crystals for use as solid biomaterials. Biomater. Sci. 2022, 10, 354–367. 10.1039/d1bm01752g. - DOI - PubMed
    1. Brodin J. D.; Ambroggio X. I.; Tang C.; Parent K. N.; Baker T. S.; Tezcan F. A. Metal-directed, chemically tunable assembly of one-, two- and three-dimensional crystalline protein arrays. Nat. Chem. 2012, 4, 375–382. 10.1038/nchem.1290. - DOI - PMC - PubMed
    1. Sontz P. A.; Bailey J. B.; Ahn S.; Tezcan F. A. A metal organic framework with spherical protein nodes: rational chemical design of 3D protein crystals. J. Am. Chem. Soc. 2015, 137, 11598–11601. 10.1021/jacs.5b07463. - DOI - PubMed
    1. Mac Sweeney A.; Chambovey A.; Wicki M.; Müller M.; Artico N.; Lange R.; Bijelic A.; Breibeck J.; Rompel A. The crystallization additive hexatungstotellurate promotes the crystallization of the HSP70 nucleotide binding domain into two different crystal forms. PLoS One 2018, 13, e019963910.1371/journal.pone.0199639. - DOI - PMC - PubMed