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Title: Crystal Structure of the alpha6beta6 Holoenzyme of propionyl-coenzyme A Carboxylase

Abstract

Propionyl-coenzyme A carboxylase (PCC), a mitochondrial biotin-dependent enzyme, is essential for the catabolism of the amino acids Thr, Val, Ile and Met, cholesterol and fatty acids with an odd number of carbon atoms. Deficiencies in PCC activity in humans are linked to the disease propionic acidaemia, an autosomal recessive disorder that can be fatal in infants. The holoenzyme of PCC is an {alpha}{sub 6}{beta}{sub 6} dodecamer, with a molecular mass of 750 kDa. The {alpha}-subunit contains the biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) domains, whereas the {beta}-subunit supplies the carboxyltransferase (CT) activity. Here we report the crystal structure at 3.2-{angstrom} resolution of a bacterial PCC {alpha}{sub 6}{beta}{sub 6} holoenzyme as well as cryo-electron microscopy (cryo-EM) reconstruction at 15-{angstrom} resolution demonstrating a similar structure for human PCC. The structure defines the overall architecture of PCC and reveals unexpectedly that the {alpha}-subunits are arranged as monomers in the holoenzyme, decorating a central {beta}{sub 6} hexamer. A hitherto unrecognized domain in the {alpha}-subunit, formed by residues between the BC and BCCP domains, is crucial for interactions with the {beta}-subunit. We have named it the BT domain. The structure reveals for the first time the relative positions of the BC andmore » CT active sites in the holoenzyme. They are separated by approximately 55 {angstrom}, indicating that the entire BCCP domain must translocate during catalysis. The BCCP domain is located in the active site of the {beta}-subunit in the current structure, providing insight for its involvement in the CT reaction. The structural information establishes a molecular basis for understanding the large collection of disease-causing mutations in PCC and is relevant for the holoenzymes of other biotin-dependent carboxylases, including 3-methylcrotonyl-CoA carboxylase (MCC) and eukaryotic acetyl-CoA carboxylase (ACC).« less

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
DOE - OFFICE OF SCIENCE
OSTI Identifier:
1019625
Report Number(s):
BNL-95470-2011-JA
Journal ID: ISSN 0028-0836; TRN: US201115%%265
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Nature
Additional Journal Information:
Journal Volume: 466; Journal Issue: 7309; Journal ID: ISSN 0028-0836
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; AMINO ACIDS; ARCHITECTURE; ATOMS; BIOCHEMISTRY; BIOLOGY; BIOTIN; CARBON; CARBOXYLASE; CARBOXYLIC ACIDS; CATABOLISM; CATALYSIS; CHOLESTEROL; CRYSTAL STRUCTURE; DISEASES; MICROSCOPY; MONOMERS; MUTATIONS; PROTEINS; RESIDUES; RESOLUTION; national synchrotron light source

Citation Formats

Huang, C, Sadre-Bazzaz, K, Shen, Y, Deng, B, Zhou, Z, and Tong, L. Crystal Structure of the alpha6beta6 Holoenzyme of propionyl-coenzyme A Carboxylase. United States: N. p., 2010. Web. doi:10.1038/nature09302.
Huang, C, Sadre-Bazzaz, K, Shen, Y, Deng, B, Zhou, Z, & Tong, L. Crystal Structure of the alpha6beta6 Holoenzyme of propionyl-coenzyme A Carboxylase. United States. doi:10.1038/nature09302.
Huang, C, Sadre-Bazzaz, K, Shen, Y, Deng, B, Zhou, Z, and Tong, L. Fri . "Crystal Structure of the alpha6beta6 Holoenzyme of propionyl-coenzyme A Carboxylase". United States. doi:10.1038/nature09302.
@article{osti_1019625,
title = {Crystal Structure of the alpha6beta6 Holoenzyme of propionyl-coenzyme A Carboxylase},
author = {Huang, C and Sadre-Bazzaz, K and Shen, Y and Deng, B and Zhou, Z and Tong, L},
abstractNote = {Propionyl-coenzyme A carboxylase (PCC), a mitochondrial biotin-dependent enzyme, is essential for the catabolism of the amino acids Thr, Val, Ile and Met, cholesterol and fatty acids with an odd number of carbon atoms. Deficiencies in PCC activity in humans are linked to the disease propionic acidaemia, an autosomal recessive disorder that can be fatal in infants. The holoenzyme of PCC is an {alpha}{sub 6}{beta}{sub 6} dodecamer, with a molecular mass of 750 kDa. The {alpha}-subunit contains the biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) domains, whereas the {beta}-subunit supplies the carboxyltransferase (CT) activity. Here we report the crystal structure at 3.2-{angstrom} resolution of a bacterial PCC {alpha}{sub 6}{beta}{sub 6} holoenzyme as well as cryo-electron microscopy (cryo-EM) reconstruction at 15-{angstrom} resolution demonstrating a similar structure for human PCC. The structure defines the overall architecture of PCC and reveals unexpectedly that the {alpha}-subunits are arranged as monomers in the holoenzyme, decorating a central {beta}{sub 6} hexamer. A hitherto unrecognized domain in the {alpha}-subunit, formed by residues between the BC and BCCP domains, is crucial for interactions with the {beta}-subunit. We have named it the BT domain. The structure reveals for the first time the relative positions of the BC and CT active sites in the holoenzyme. They are separated by approximately 55 {angstrom}, indicating that the entire BCCP domain must translocate during catalysis. The BCCP domain is located in the active site of the {beta}-subunit in the current structure, providing insight for its involvement in the CT reaction. The structural information establishes a molecular basis for understanding the large collection of disease-causing mutations in PCC and is relevant for the holoenzymes of other biotin-dependent carboxylases, including 3-methylcrotonyl-CoA carboxylase (MCC) and eukaryotic acetyl-CoA carboxylase (ACC).},
doi = {10.1038/nature09302},
journal = {Nature},
issn = {0028-0836},
number = 7309,
volume = 466,
place = {United States},
year = {2010},
month = {1}
}