Cryo-EM Analysis Reveals New Insights into the Mechanism of Action of Pyruvate Carboxylase

Gorka Lasso; Linda P.C. Yu; David Gil; Song Xiang; Liang Tong; Mikel Valle

doi:10.1016/j.str.2010.07.008

Cryo-EM Analysis Reveals New Insights into the Mechanism of Action of Pyruvate Carboxylase

Gorka Lasso, Linda P.C. Yu, David Gil, Song Xiang, Liang Tong, Mikel Valle

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

Pyruvate carboxylase (PC) is a conserved multifunctional enzyme linked to important metabolic diseases. PC homotetramer is arranged in two layers with two opposing monomers per layer. Cryo-EM explores the conformational variability of PC in the presence of different substrates. The results demonstrate that the biotin-carboxyl carrier protein (BCCP) domain localizes near the biotin carboxylase (BC) domain of its own monomer and travels to the carboxyltransferase (CT) domain of the opposite monomer. All density maps show noticeable conformational differences between layers, mainly for the BCCP and BC domains. This asymmetry may be indicative of a coordination mechanism where monomers from different layers catalyze the BC and CT reactions consecutively. A conformational change of the PC tetramerization (PT) domain suggests a new functional role in communication. A long-range communication pathway between subunits in different layers, via interacting PT-PT and BC-BC domains, may be responsible for the cooperativity of PC from Staphylococcus aureus.

Original language	English (US)
Pages (from-to)	1300-1310
Number of pages	11
Journal	Structure
Volume	18
Issue number	10
DOIs	https://doi.org/10.1016/j.str.2010.07.008
State	Published - Oct 13 2010
Externally published	Yes

ASJC Scopus subject areas

Structural Biology
Molecular Biology

Access to Document

10.1016/j.str.2010.07.008

Cite this

@article{28c44cd6be8c4d1d8585e1ae5b83d271,

title = "Cryo-EM Analysis Reveals New Insights into the Mechanism of Action of Pyruvate Carboxylase",

abstract = "Pyruvate carboxylase (PC) is a conserved multifunctional enzyme linked to important metabolic diseases. PC homotetramer is arranged in two layers with two opposing monomers per layer. Cryo-EM explores the conformational variability of PC in the presence of different substrates. The results demonstrate that the biotin-carboxyl carrier protein (BCCP) domain localizes near the biotin carboxylase (BC) domain of its own monomer and travels to the carboxyltransferase (CT) domain of the opposite monomer. All density maps show noticeable conformational differences between layers, mainly for the BCCP and BC domains. This asymmetry may be indicative of a coordination mechanism where monomers from different layers catalyze the BC and CT reactions consecutively. A conformational change of the PC tetramerization (PT) domain suggests a new functional role in communication. A long-range communication pathway between subunits in different layers, via interacting PT-PT and BC-BC domains, may be responsible for the cooperativity of PC from Staphylococcus aureus.",

author = "Gorka Lasso and Yu, {Linda P.C.} and David Gil and Song Xiang and Liang Tong and Mikel Valle",

note = "Funding Information: We thank Melisa L{\'a}zaro for assistance during cryo-EM image processing. This work was supported by the Etortek Research Programmes 2008/2010 (Department of Industry, Tourism and Trade of the Government of the Autonomous Community of the Basque Country) (M.V.), the Innovation Technology Department of the Bizkaia County (M.V.), the National Institutes of Health (DK067238 to L.T.), and Caja Navarra (16507). L.P.C.Y. is also supported by a training program in Cellular and Molecular Foundations of Biomedical Science from the NIH (GM008798). ",

year = "2010",

month = oct,

day = "13",

doi = "10.1016/j.str.2010.07.008",

language = "English (US)",

volume = "18",

pages = "1300--1310",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "10",

}

TY - JOUR

T1 - Cryo-EM Analysis Reveals New Insights into the Mechanism of Action of Pyruvate Carboxylase

AU - Lasso, Gorka

AU - Yu, Linda P.C.

AU - Gil, David

AU - Xiang, Song

AU - Tong, Liang

AU - Valle, Mikel

N1 - Funding Information: We thank Melisa Lázaro for assistance during cryo-EM image processing. This work was supported by the Etortek Research Programmes 2008/2010 (Department of Industry, Tourism and Trade of the Government of the Autonomous Community of the Basque Country) (M.V.), the Innovation Technology Department of the Bizkaia County (M.V.), the National Institutes of Health (DK067238 to L.T.), and Caja Navarra (16507). L.P.C.Y. is also supported by a training program in Cellular and Molecular Foundations of Biomedical Science from the NIH (GM008798).

PY - 2010/10/13

Y1 - 2010/10/13

N2 - Pyruvate carboxylase (PC) is a conserved multifunctional enzyme linked to important metabolic diseases. PC homotetramer is arranged in two layers with two opposing monomers per layer. Cryo-EM explores the conformational variability of PC in the presence of different substrates. The results demonstrate that the biotin-carboxyl carrier protein (BCCP) domain localizes near the biotin carboxylase (BC) domain of its own monomer and travels to the carboxyltransferase (CT) domain of the opposite monomer. All density maps show noticeable conformational differences between layers, mainly for the BCCP and BC domains. This asymmetry may be indicative of a coordination mechanism where monomers from different layers catalyze the BC and CT reactions consecutively. A conformational change of the PC tetramerization (PT) domain suggests a new functional role in communication. A long-range communication pathway between subunits in different layers, via interacting PT-PT and BC-BC domains, may be responsible for the cooperativity of PC from Staphylococcus aureus.

AB - Pyruvate carboxylase (PC) is a conserved multifunctional enzyme linked to important metabolic diseases. PC homotetramer is arranged in two layers with two opposing monomers per layer. Cryo-EM explores the conformational variability of PC in the presence of different substrates. The results demonstrate that the biotin-carboxyl carrier protein (BCCP) domain localizes near the biotin carboxylase (BC) domain of its own monomer and travels to the carboxyltransferase (CT) domain of the opposite monomer. All density maps show noticeable conformational differences between layers, mainly for the BCCP and BC domains. This asymmetry may be indicative of a coordination mechanism where monomers from different layers catalyze the BC and CT reactions consecutively. A conformational change of the PC tetramerization (PT) domain suggests a new functional role in communication. A long-range communication pathway between subunits in different layers, via interacting PT-PT and BC-BC domains, may be responsible for the cooperativity of PC from Staphylococcus aureus.

UR - http://www.scopus.com/inward/record.url?scp=77957756727&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957756727&partnerID=8YFLogxK

U2 - 10.1016/j.str.2010.07.008

DO - 10.1016/j.str.2010.07.008

M3 - Article

C2 - 20947019

AN - SCOPUS:77957756727

SN - 0969-2126

VL - 18

SP - 1300

EP - 1310

JO - Structure

JF - Structure

IS - 10

ER -

Cryo-EM Analysis Reveals New Insights into the Mechanism of Action of Pyruvate Carboxylase

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this