7NS9

Triphosphate tunnel metalloenzyme from Sulfolobus acidocaldarius in complex with triphosphate and calcium

Summary for 7NS9

• Entry DOI: 10.2210/pdb7ns9/pdb
• Descriptor: Triphosphate tunnel metalloenzyme Saci_0718, ACETATE ION, CALCIUM ION, ... (6 entities in total)
• Functional Keywords: ttm, cyth, triphosphate, calcium, hydrolase
• Biological source: Sulfolobus acidocaldarius (strain ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770)
• Total number of polymer chains: 1
• Total formula weight: 24202.25

Authors

Vogt, M.S.,Essen, L.-O.,Banerjee, A. (deposition date: 2021-03-05, release date: 2021-06-02, Last modification date: 2024-01-31)

Primary citation

Vogt, M.S.,Ngouoko Nguepbeu, R.R.,Mohr, M.K.F.,Albers, S.V.,Essen, L.O.,Banerjee, A.
The archaeal triphosphate tunnel metalloenzyme SaTTM defines structural determinants for the diverse activities in the CYTH protein family.
J.Biol.Chem., 297:100820-100820, 2021

PubMed Abstract: CYTH proteins make up a large superfamily that is conserved in all three domains of life. These enzymes have a triphosphate tunnel metalloenzyme (TTM) fold, which typically results in phosphatase functions, e.g., RNA triphosphatase, inorganic polyphosphatase, or thiamine triphosphatase. Some CYTH orthologs cyclize nucleotide triphosphates to 3',5'-cyclic nucleotides. So far, archaeal CYTH proteins have been annotated as adenylyl cyclases, although experimental evidence to support these annotations is lacking. To address this gap, we characterized a CYTH ortholog, SaTTM, from the crenarchaeote Sulfolobus acidocaldarius. Our in silico studies derived ten major subclasses within the CYTH family implying a close relationship between these archaeal CYTH enzymes and class IV adenylyl cyclases. However, initial biochemical characterization reveals inability of SaTTM to produce any cyclic nucleotides. Instead, our structural and functional analyses show a classical TTM behavior, i.e., triphosphatase activity, where pyrophosphate causes product inhibition. The Ca-inhibited Michaelis complex indicates a two-metal-ion reaction mechanism analogous to other TTMs. Cocrystal structures of SaTTM further reveal conformational dynamics in SaTTM that suggest feedback inhibition in TTMs due to tunnel closure in the product state. These structural insights combined with further sequence similarity network-based in silico analyses provide a firm molecular basis for distinguishing CYTH orthologs with phosphatase activities from class IV adenylyl cyclases.

PubMed: 34029589
DOI: 10.1016/j.jbc.2021.100820

Experimental method

X-RAY DIFFRACTION (1.75 Å)