Structure and Mechanism of Acetylation by the N-Terminal Dual Enzyme NatA/Naa50 Complex

Sunbin Deng; Robert S Magin; Xuepeng Wei; Buyan Pan; E James Petersson; Ronen Marmorstein

doi:10.1016/j.str.2019.04.014

Structure and Mechanism of Acetylation by the N-Terminal Dual Enzyme NatA/Naa50 Complex

Structure. 2019 Jul 2;27(7):1057-1070.e4. doi: 10.1016/j.str.2019.04.014. Epub 2019 May 30.

Authors

Sunbin Deng¹, Robert S Magin², Xuepeng Wei³, Buyan Pan⁴, E James Petersson⁵, Ronen Marmorstein⁶

Affiliations

¹ Department of Chemistry, University of Pennsylvania, 231 South 34(th) Street, Philadelphia, PA 19104, USA; Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
² Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
³ Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biochemistry and Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, 421 Curie Boulevard, Philadelphia, PA 19104, USA.
⁴ Department of Chemistry, University of Pennsylvania, 231 South 34(th) Street, Philadelphia, PA 19104, USA.
⁵ Department of Chemistry, University of Pennsylvania, 231 South 34(th) Street, Philadelphia, PA 19104, USA; Department of Biochemistry and Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, 421 Curie Boulevard, Philadelphia, PA 19104, USA.
⁶ Department of Chemistry, University of Pennsylvania, 231 South 34(th) Street, Philadelphia, PA 19104, USA; Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biochemistry and Biophysics, Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, 421 Curie Boulevard, Philadelphia, PA 19104, USA. Electronic address: [email protected].

Abstract

NatA co-translationally acetylates the N termini of over 40% of eukaryotic proteins and can associate with another catalytic subunit, Naa50, to form a ternary NatA/Naa50 dual enzyme complex (also called NatE). The molecular basis of association between Naa50 and NatA and the mechanism for how their association affects their catalytic activities in yeast and human are poorly understood. Here, we determined the X-ray crystal structure of yeast NatA/Naa50 as a scaffold to understand coregulation of NatA/Naa50 activity in both yeast and human. We find that Naa50 makes evolutionarily conserved contacts to both the Naa10 and Naa15 subunits of NatA. These interactions promote catalytic crosstalk within the human complex, but do so to a lesser extent in the yeast complex, where Naa50 activity is compromised. These studies have implications for understanding the role of the NatA/Naa50 complex in modulating the majority of the N-terminal acetylome in diverse species.

Keywords: N-terminal acetylation; Naa50; NatA; NatE; X-ray crystallography; protein complex.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Acetyl Coenzyme A / chemistry
Acetyl Coenzyme A / metabolism
Acetylation
Acetyltransferases / chemistry*
Acetyltransferases / genetics
Acetyltransferases / metabolism
Animals
Catalytic Domain
Cloning, Molecular
Crystallography, X-Ray
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Humans
Isoenzymes / chemistry
Isoenzymes / genetics
Isoenzymes / metabolism
Kinetics
Models, Molecular
Multienzyme Complexes / chemistry*
Multienzyme Complexes / genetics
Multienzyme Complexes / metabolism
N-Terminal Acetyltransferase A / chemistry*
N-Terminal Acetyltransferase A / genetics
N-Terminal Acetyltransferase A / metabolism
N-Terminal Acetyltransferase E / chemistry*
N-Terminal Acetyltransferase E / genetics
N-Terminal Acetyltransferase E / metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Protein Processing, Post-Translational*
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Saccharomyces cerevisiae / enzymology*
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae Proteins / chemistry*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism
Sf9 Cells
Spodoptera
Substrate Specificity

Substances

Isoenzymes
Multienzyme Complexes
NAA15 protein, human
Recombinant Proteins
Saccharomyces cerevisiae Proteins
Acetyl Coenzyme A
Acetyltransferases
NatA protein, S cerevisiae
N-Terminal Acetyltransferase A
NAA10 protein, human
N-Terminal Acetyltransferase E
NAA50 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding