Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers

UDP-Glc:protein transglucosylase (UPTG) [EC 2.4.1.112], the protein regarded as pivotal to starch biosynthesis initiation, was purified from potato (Solanum tuberosum) tubers by modification of previous procedures. The steps used for purification were: solubilization of the particulate potato tuber...

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Detalles Bibliográficos
Publicado: 1997
Materias:
Enzyme purification
potato tuber
Solanum tuberosum
starch synthesis initiation
UDP-glucose:protein transglucosylase
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09819428_v35_n3_p205_Bocca
http://hdl.handle.net/20.500.12110/paper_09819428_v35_n3_p205_Bocca
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paper:paper_09819428_v35_n3_p205_Bocca
record_format dspace
spelling paper:paper_09819428_v35_n3_p205_Bocca2023-06-08T15:59:13Z Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers Enzyme purification potato tuber Solanum tuberosum starch synthesis initiation UDP-glucose:protein transglucosylase UDP-Glc:protein transglucosylase (UPTG) [EC 2.4.1.112], the protein regarded as pivotal to starch biosynthesis initiation, was purified from potato (Solanum tuberosum) tubers by modification of previous procedures. The steps used for purification were: solubilization of the particulate potato tuber preparation, ion-exchange chromatographies on DEAE-Sepharose fast flow and Mono Q columns, and affinity chromatography on a UDP-hexanolamine-agarose column. The last step yielded an electrophoretically homogeneous, stable enzyme protein with 1063-fold purification and a reproducible yield of 27%. UPTG, which undergoes self-glucosylation in an UDP-Glc and Mn2+-dependent reaction, is homomeric, made up of 38-kDa subunits. Polyclonal antibodies raised in rabbits against the potato tuber 38-kDa polypeptide cross reacted strongly with the potato tuber UPTG and with the maize (Zea mays) endosperm enzyme as well, and neutralized (inhibited) potato tuber and maize endosperm UPTG activity (more than 90%). This is the first demonstration that highly specific antibodies raised against one single and even denatured-protein are capable of neutralizing UPTG activity. The availability of the purified protein and specific antibodies opens the way to molecular characterization of potato tuber UPTG and to studying its role in the initiation of starch synthesis. 1997 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09819428_v35_n3_p205_Bocca http://hdl.handle.net/20.500.12110/paper_09819428_v35_n3_p205_Bocca
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Enzyme purification
potato tuber
Solanum tuberosum
starch synthesis initiation
UDP-glucose:protein transglucosylase
spellingShingle Enzyme purification
potato tuber
Solanum tuberosum
starch synthesis initiation
UDP-glucose:protein transglucosylase
Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
topic_facet Enzyme purification
potato tuber
Solanum tuberosum
starch synthesis initiation
UDP-glucose:protein transglucosylase
description UDP-Glc:protein transglucosylase (UPTG) [EC 2.4.1.112], the protein regarded as pivotal to starch biosynthesis initiation, was purified from potato (Solanum tuberosum) tubers by modification of previous procedures. The steps used for purification were: solubilization of the particulate potato tuber preparation, ion-exchange chromatographies on DEAE-Sepharose fast flow and Mono Q columns, and affinity chromatography on a UDP-hexanolamine-agarose column. The last step yielded an electrophoretically homogeneous, stable enzyme protein with 1063-fold purification and a reproducible yield of 27%. UPTG, which undergoes self-glucosylation in an UDP-Glc and Mn2+-dependent reaction, is homomeric, made up of 38-kDa subunits. Polyclonal antibodies raised in rabbits against the potato tuber 38-kDa polypeptide cross reacted strongly with the potato tuber UPTG and with the maize (Zea mays) endosperm enzyme as well, and neutralized (inhibited) potato tuber and maize endosperm UPTG activity (more than 90%). This is the first demonstration that highly specific antibodies raised against one single and even denatured-protein are capable of neutralizing UPTG activity. The availability of the purified protein and specific antibodies opens the way to molecular characterization of potato tuber UPTG and to studying its role in the initiation of starch synthesis.
title Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
title_short Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
title_full Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
title_fullStr Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
title_full_unstemmed Initiation of starch biosynthesis: Purification and characterization of UDP-glucose:protein transglucosylase from potato tubers
title_sort initiation of starch biosynthesis: purification and characterization of udp-glucose:protein transglucosylase from potato tubers
publishDate 1997
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09819428_v35_n3_p205_Bocca
http://hdl.handle.net/20.500.12110/paper_09819428_v35_n3_p205_Bocca
_version_ 1768546594979840000

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