The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms

Misty L. Kuhn; Carlos M. Figueroa; Alberto A. Iglesias; Miguel Ballicora

doi:10.1186/1471-2148-13-51

The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms

Misty L. Kuhn, Carlos M. Figueroa, Alberto A. Iglesias, Miguel Ballicora

Department of Chemistry and Biochemistry

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

Abstract

<p> <h4> Background </h4></p><p> ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in the synthesis of glycogen in bacteria and starch in algae and plants. In oxygenic photosynthetic organisms, ADP-Glc PPase is mainly activated by 3-phosphoglycerate (3-PGA) and to a lesser extent by other metabolites. In this work, we analyzed the activation promiscuity of ADP-Glc PPase subunits from the cyanobacterium <em> Anabaena </em> PCC 7120, the green alga <em> Ostreococcus tauri </em> , and potato ( <em> Solanum tuberosum </em> ) tuber by comparing a specificity constant for 3-PGA, fructose-1,6-bisphosphate (FBP), fructose-6-phosphate, and glucose-6-phosphate. <h4> Results </h4></p><p> The 3-PGA specificity constant for the enzymes from <em> Anabaena </em> (homotetramer), <em> O. tauri </em> , and potato tuber was considerably higher than for other activators. <em> O. tauri </em> and potato tuber enzymes were heterotetramers comprising homologous small and large subunits. Conversely, the <em> O. tauri </em> small subunit (OtaS) homotetramer was more promiscuous because its FBP specificity constant was similar to that for 3-PGA. To explore the role of both OtaS and OtaL ( <em> O. tauri </em> large subunit) in determining the specificity of the heterotetramer, we knocked out the catalytic activity of each subunit individually by site-directed mutagenesis. Interestingly, the mutants OtaS <sub> D148A </sub> /OtaL and OtaS/OtaL <sub> D171A </sub> had higher specificity constants for 3-PGA than for FBP. <h4> Conclusions </h4></p><p> After gene duplication, OtaS seemed to have lost specificity for 3-PGA compared to FBP. This was physiologically and evolutionarily feasible because co-expression of both subunits restored the specificity for 3-PGA of the resulting heterotetrameric wild type enzyme. This widespread promiscuity seems to be ancestral and intrinsic to the enzyme family. Its presence could constitute an efficient evolutionary mechanism to accommodate the ADP-Glc PPase regulation to different metabolic needs.</p>

Original language	American English
Journal	Chemistry: Faculty Publications and Other Works
Volume	13
Issue number	51
DOIs	https://doi.org/10.1186/1471-2148-13-51
State	Published - Feb 21 2013

Keywords

evolutionary biology
ADP-glucose pyrophosphorylases
oxygenic photosynthetic organisms

Disciplines

Chemistry

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@article{809a3c3449194b77adaf7525719d4447,

title = "The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms",

abstract = " Background ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in the synthesis of glycogen in bacteria and starch in algae and plants. In oxygenic photosynthetic organisms, ADP-Glc PPase is mainly activated by 3-phosphoglycerate (3-PGA) and to a lesser extent by other metabolites. In this work, we analyzed the activation promiscuity of ADP-Glc PPase subunits from the cyanobacterium Anabaena PCC 7120, the green alga Ostreococcus tauri , and potato ( Solanum tuberosum ) tuber by comparing a specificity constant for 3-PGA, fructose-1,6-bisphosphate (FBP), fructose-6-phosphate, and glucose-6-phosphate. Results The 3-PGA specificity constant for the enzymes from Anabaena (homotetramer), O. tauri , and potato tuber was considerably higher than for other activators. O. tauri and potato tuber enzymes were heterotetramers comprising homologous small and large subunits. Conversely, the O. tauri small subunit (OtaS) homotetramer was more promiscuous because its FBP specificity constant was similar to that for 3-PGA. To explore the role of both OtaS and OtaL ( O. tauri large subunit) in determining the specificity of the heterotetramer, we knocked out the catalytic activity of each subunit individually by site-directed mutagenesis. Interestingly, the mutants OtaS D148A /OtaL and OtaS/OtaL D171A had higher specificity constants for 3-PGA than for FBP. Conclusions After gene duplication, OtaS seemed to have lost specificity for 3-PGA compared to FBP. This was physiologically and evolutionarily feasible because co-expression of both subunits restored the specificity for 3-PGA of the resulting heterotetrameric wild type enzyme. This widespread promiscuity seems to be ancestral and intrinsic to the enzyme family. Its presence could constitute an efficient evolutionary mechanism to accommodate the ADP-Glc PPase regulation to different metabolic needs.",

keywords = "evolutionary biology, ADP-glucose pyrophosphorylases, oxygenic photosynthetic organisms",

author = "Kuhn, {Misty L.} and Figueroa, {Carlos M.} and Iglesias, {Alberto A.} and Miguel Ballicora",

note = "Kuhn, Misty L., Carlos M. Figueroa, Alberto A. Iglesias, and Miguel A. Ballicora. {"}The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms.{"} BMC Evolutionary Biology, Volume 13, Issue 51, February 21, 2013, http://dx.doi.org/10.1186/1471-2148-13-51.",

year = "2013",

month = feb,

day = "21",

doi = "10.1186/1471-2148-13-51",

language = "American English",

volume = "13",

journal = "Chemistry: Faculty Publications and Other Works",

number = "51",

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TY - JOUR

T1 - The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms

AU - Kuhn, Misty L.

AU - Figueroa, Carlos M.

AU - Iglesias, Alberto A.

AU - Ballicora, Miguel

N1 - Kuhn, Misty L., Carlos M. Figueroa, Alberto A. Iglesias, and Miguel A. Ballicora. "The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms." BMC Evolutionary Biology, Volume 13, Issue 51, February 21, 2013, http://dx.doi.org/10.1186/1471-2148-13-51.

PY - 2013/2/21

Y1 - 2013/2/21

N2 - Background ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in the synthesis of glycogen in bacteria and starch in algae and plants. In oxygenic photosynthetic organisms, ADP-Glc PPase is mainly activated by 3-phosphoglycerate (3-PGA) and to a lesser extent by other metabolites. In this work, we analyzed the activation promiscuity of ADP-Glc PPase subunits from the cyanobacterium Anabaena PCC 7120, the green alga Ostreococcus tauri , and potato ( Solanum tuberosum ) tuber by comparing a specificity constant for 3-PGA, fructose-1,6-bisphosphate (FBP), fructose-6-phosphate, and glucose-6-phosphate. Results The 3-PGA specificity constant for the enzymes from Anabaena (homotetramer), O. tauri , and potato tuber was considerably higher than for other activators. O. tauri and potato tuber enzymes were heterotetramers comprising homologous small and large subunits. Conversely, the O. tauri small subunit (OtaS) homotetramer was more promiscuous because its FBP specificity constant was similar to that for 3-PGA. To explore the role of both OtaS and OtaL ( O. tauri large subunit) in determining the specificity of the heterotetramer, we knocked out the catalytic activity of each subunit individually by site-directed mutagenesis. Interestingly, the mutants OtaS D148A /OtaL and OtaS/OtaL D171A had higher specificity constants for 3-PGA than for FBP. Conclusions After gene duplication, OtaS seemed to have lost specificity for 3-PGA compared to FBP. This was physiologically and evolutionarily feasible because co-expression of both subunits restored the specificity for 3-PGA of the resulting heterotetrameric wild type enzyme. This widespread promiscuity seems to be ancestral and intrinsic to the enzyme family. Its presence could constitute an efficient evolutionary mechanism to accommodate the ADP-Glc PPase regulation to different metabolic needs.

AB - Background ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in the synthesis of glycogen in bacteria and starch in algae and plants. In oxygenic photosynthetic organisms, ADP-Glc PPase is mainly activated by 3-phosphoglycerate (3-PGA) and to a lesser extent by other metabolites. In this work, we analyzed the activation promiscuity of ADP-Glc PPase subunits from the cyanobacterium Anabaena PCC 7120, the green alga Ostreococcus tauri , and potato ( Solanum tuberosum ) tuber by comparing a specificity constant for 3-PGA, fructose-1,6-bisphosphate (FBP), fructose-6-phosphate, and glucose-6-phosphate. Results The 3-PGA specificity constant for the enzymes from Anabaena (homotetramer), O. tauri , and potato tuber was considerably higher than for other activators. O. tauri and potato tuber enzymes were heterotetramers comprising homologous small and large subunits. Conversely, the O. tauri small subunit (OtaS) homotetramer was more promiscuous because its FBP specificity constant was similar to that for 3-PGA. To explore the role of both OtaS and OtaL ( O. tauri large subunit) in determining the specificity of the heterotetramer, we knocked out the catalytic activity of each subunit individually by site-directed mutagenesis. Interestingly, the mutants OtaS D148A /OtaL and OtaS/OtaL D171A had higher specificity constants for 3-PGA than for FBP. Conclusions After gene duplication, OtaS seemed to have lost specificity for 3-PGA compared to FBP. This was physiologically and evolutionarily feasible because co-expression of both subunits restored the specificity for 3-PGA of the resulting heterotetrameric wild type enzyme. This widespread promiscuity seems to be ancestral and intrinsic to the enzyme family. Its presence could constitute an efficient evolutionary mechanism to accommodate the ADP-Glc PPase regulation to different metabolic needs.

KW - evolutionary biology

KW - ADP-glucose pyrophosphorylases

KW - oxygenic photosynthetic organisms

UR - https://ecommons.luc.edu/chemistry_facpubs/44

U2 - 10.1186/1471-2148-13-51

DO - 10.1186/1471-2148-13-51

M3 - Article

VL - 13

JO - Chemistry: Faculty Publications and Other Works

JF - Chemistry: Faculty Publications and Other Works

IS - 51

ER -

The Ancestral Activation Promiscuity of ADP-glucose Pyrophosphorylases from Oxygenic Photosynthetic Organisms

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