Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers

Simon Hald, Andreas Blennow, Allan Stensballe, Guy Bauw, Karen Gjesing Welinder

Research output: Contribution to conference without publisher/journalPosterResearch

Abstract

In potato (Solanum tuberosum) tuber starch is synthesized and stored in amyloplasts. Amyloplasts were prepared from in vitro or agar-grown micro tubers and from soil-grown mini tubers and subjected to proteome analysis. The quality of amyloplasts was assessed by comparing amyloplast fractions and total tuber extracts by SDS-PAGE and the specific activities of marker enzymes for amyloplast, cytosol, mitochondria and the vacuole. SDS-PAGE separated amyloplast and starch granule proteins were in-gel digested with trypsin, analyzed by mass spectrometry, and identified by searches against presently available potato protein sequences. Some of these proteins were demonstrated to localize to the amyloplast stroma for the first time. The micro and mini tuber proteomes were very different. However, starch phosphorylase L-1 was particularly abundant in both proteomes. Moreover, disproportionating enzyme 1 and 2, starch phosphorylase L-2 and H, and a-glucan water dikinase were found within the amyloplast, supporting that these specific starch degradative reactions occur within intact potato tuber plastids independently of potential loss of amyloplast envelope integrity during sprouting. The majority of the identified proteins had a predicted plastid transit peptide supporting their presence in the amyloplast. Remarkably, catechol oxidase and transketolase showed twin-arginine translocation (Tat) motives, and 'putative' deoxynucleoside kinase a Sec secretory motif targeting for thylakoid or thylakoid-like structures not expected in amyloplast.
Original languageEnglish
Publication date2006
Number of pages1
Publication statusPublished - 2006
Event7th Siena Meeting. From genome to proteome: Back to the future - Siena, Italy
Duration: 3 Sep 20067 Sep 2006

Conference

Conference7th Siena Meeting. From genome to proteome: Back to the future
CountryItaly
CitySiena
Period03/09/200607/09/2006

Fingerprint

amyloplasts
tubers
agar
potatoes
soil
proteins
proteome
starch
phosphorylase
thylakoids
plastids
potato protein
transketolase
glucans
sprouting
starch granules
catechol oxidase
cytosol
Solanum tuberosum
trypsin

Cite this

Hald, S., Blennow, A., Stensballe, A., Bauw, G., & Welinder, K. G. (2006). Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers. Poster session presented at 7th Siena Meeting. From genome to proteome: Back to the future, Siena, Italy.
Hald, Simon ; Blennow, Andreas ; Stensballe, Allan ; Bauw, Guy ; Welinder, Karen Gjesing. / Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers. Poster session presented at 7th Siena Meeting. From genome to proteome: Back to the future, Siena, Italy.1 p.
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abstract = "In potato (Solanum tuberosum) tuber starch is synthesized and stored in amyloplasts. Amyloplasts were prepared from in vitro or agar-grown micro tubers and from soil-grown mini tubers and subjected to proteome analysis. The quality of amyloplasts was assessed by comparing amyloplast fractions and total tuber extracts by SDS-PAGE and the specific activities of marker enzymes for amyloplast, cytosol, mitochondria and the vacuole. SDS-PAGE separated amyloplast and starch granule proteins were in-gel digested with trypsin, analyzed by mass spectrometry, and identified by searches against presently available potato protein sequences. Some of these proteins were demonstrated to localize to the amyloplast stroma for the first time. The micro and mini tuber proteomes were very different. However, starch phosphorylase L-1 was particularly abundant in both proteomes. Moreover, disproportionating enzyme 1 and 2, starch phosphorylase L-2 and H, and a-glucan water dikinase were found within the amyloplast, supporting that these specific starch degradative reactions occur within intact potato tuber plastids independently of potential loss of amyloplast envelope integrity during sprouting. The majority of the identified proteins had a predicted plastid transit peptide supporting their presence in the amyloplast. Remarkably, catechol oxidase and transketolase showed twin-arginine translocation (Tat) motives, and 'putative' deoxynucleoside kinase a Sec secretory motif targeting for thylakoid or thylakoid-like structures not expected in amyloplast.",
author = "Simon Hald and Andreas Blennow and Allan Stensballe and Guy Bauw and Welinder, {Karen Gjesing}",
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language = "English",
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Hald, S, Blennow, A, Stensballe, A, Bauw, G & Welinder, KG 2006, 'Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers', Siena, Italy, 03/09/2006 - 07/09/2006, .

Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers. / Hald, Simon; Blennow, Andreas; Stensballe, Allan; Bauw, Guy; Welinder, Karen Gjesing.

2006. Poster session presented at 7th Siena Meeting. From genome to proteome: Back to the future, Siena, Italy.

Research output: Contribution to conference without publisher/journalPosterResearch

TY - CONF

T1 - Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers

AU - Hald, Simon

AU - Blennow, Andreas

AU - Stensballe, Allan

AU - Bauw, Guy

AU - Welinder, Karen Gjesing

PY - 2006

Y1 - 2006

N2 - In potato (Solanum tuberosum) tuber starch is synthesized and stored in amyloplasts. Amyloplasts were prepared from in vitro or agar-grown micro tubers and from soil-grown mini tubers and subjected to proteome analysis. The quality of amyloplasts was assessed by comparing amyloplast fractions and total tuber extracts by SDS-PAGE and the specific activities of marker enzymes for amyloplast, cytosol, mitochondria and the vacuole. SDS-PAGE separated amyloplast and starch granule proteins were in-gel digested with trypsin, analyzed by mass spectrometry, and identified by searches against presently available potato protein sequences. Some of these proteins were demonstrated to localize to the amyloplast stroma for the first time. The micro and mini tuber proteomes were very different. However, starch phosphorylase L-1 was particularly abundant in both proteomes. Moreover, disproportionating enzyme 1 and 2, starch phosphorylase L-2 and H, and a-glucan water dikinase were found within the amyloplast, supporting that these specific starch degradative reactions occur within intact potato tuber plastids independently of potential loss of amyloplast envelope integrity during sprouting. The majority of the identified proteins had a predicted plastid transit peptide supporting their presence in the amyloplast. Remarkably, catechol oxidase and transketolase showed twin-arginine translocation (Tat) motives, and 'putative' deoxynucleoside kinase a Sec secretory motif targeting for thylakoid or thylakoid-like structures not expected in amyloplast.

AB - In potato (Solanum tuberosum) tuber starch is synthesized and stored in amyloplasts. Amyloplasts were prepared from in vitro or agar-grown micro tubers and from soil-grown mini tubers and subjected to proteome analysis. The quality of amyloplasts was assessed by comparing amyloplast fractions and total tuber extracts by SDS-PAGE and the specific activities of marker enzymes for amyloplast, cytosol, mitochondria and the vacuole. SDS-PAGE separated amyloplast and starch granule proteins were in-gel digested with trypsin, analyzed by mass spectrometry, and identified by searches against presently available potato protein sequences. Some of these proteins were demonstrated to localize to the amyloplast stroma for the first time. The micro and mini tuber proteomes were very different. However, starch phosphorylase L-1 was particularly abundant in both proteomes. Moreover, disproportionating enzyme 1 and 2, starch phosphorylase L-2 and H, and a-glucan water dikinase were found within the amyloplast, supporting that these specific starch degradative reactions occur within intact potato tuber plastids independently of potential loss of amyloplast envelope integrity during sprouting. The majority of the identified proteins had a predicted plastid transit peptide supporting their presence in the amyloplast. Remarkably, catechol oxidase and transketolase showed twin-arginine translocation (Tat) motives, and 'putative' deoxynucleoside kinase a Sec secretory motif targeting for thylakoid or thylakoid-like structures not expected in amyloplast.

M3 - Poster

ER -

Hald S, Blennow A, Stensballe A, Bauw G, Welinder KG. Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers. 2006. Poster session presented at 7th Siena Meeting. From genome to proteome: Back to the future, Siena, Italy.