Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean.

Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Research Abstract Details 

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Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Abstract Text:

Van Hai Nong,Masaomi Arahira,Van Chi Phan,Chan-Shick Kim,Deyu Zhang,Kyoko Udaka,Chikafusa Fukazawa,

Molecular characterization of plant group II chaperonin (CCT, c-cpn, or TriC) still remains elusive. By PCR-based cloning techniques using soybeans, we have made a successful attempt to clone a delta-subunit homologue of CCT (CCTdelta). This subunit is responsible for the binding of an in vivo substrate, alpha-actin, by assisting the correct folding of the cytoskeletal protein in mouse, and the occurrence of the subunit homologue in plant CCT was unclear. As the cloning strategy, a putative amino acid segment, NH(2)-Gly-Gly-Gly-Ala-Pro-Glu-COOH, which is tightly conserved in all known animal and yeast CCTdelta subunits, was chosen for designing a degenerate primer of the PCR-cloning. The resultant 1881-bp cDNA was found to have an open-reading frame of 533 amino acids with a calculated molecular mass of 57,677 Da and to share about 58-65% identity overall at the amino acid level with the corresponding subunits known to date. Using antibodies raised against Escherichia coli-produced soybean insoluble CCTdelta as a monitoring tool, we purified soybean CCT from the extract of its immature seeds. STEM images demonstrated that the molecular shape of soybean CCT is a double eight-membered ring, which resembles the known group II chaperonins. The CCT also reactivated a denatured firefly luciferase with a significant, but limited level of the native enzymic activity in an in vitro system. Northern blot analysis showed that soybean CCTdelta gene, which is intronless and composed of a small family, was only expressed at a very early stage of seed development of soybean.

Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Publishing Authors By Initials

VH Nong,M Arahira,VC Phan,CS Kim,D Zhang,K Udaka,C Fukazawa,

For similar plants: plant families and groups: angiosperms: fabaceae: soybeans research abstracts see: plants: plant families and groups: angiosperms: fabaceae: soybeans research

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Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 132

Page Numbers: 291-300

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Aug

YEAR: 2002

Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Keywords Mesh Terms:

KEYWORDS: Soybeans

MESH TERMS: metabolism

Chemical & Substance for Abstract: Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean. Information

Substance Name: Luciferases

Registry Number: EC 1.13.12.-

Grant and Affiliation Information for Molecular cloning and characterization of a group II chaperonin delta-subunit from soybean.

AFFILIATION: Genetic Engineering Laboratory, National Food Research Institute, Ministry of Agriculture, Forestry And Fisheries, Tsukuba Science City, Ibaraki 305-8642, Japan.

Country: Japan

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MEDLINETA: J Biochem

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ACCESSION NUMBER: AB004233

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