tailieunhanh - Báo cáo khoa học: Molecular cloning and characterization of two soybean protein disulfide isomerases as molecular chaperones for seed storage proteins

Protein disulfide isomerase family proteins play important roles in the fold-ing of nascent polypeptides and the formation of disulfide bonds in the endoplasmic reticulum. In this study, we cloned two similar protein disul-fide isomerase family genes from soybean leaf (Glycine maxL. Merrill. cv Jack). The cDNAs encode proteins of 525 and 551 amino acids, named GmPDIL-1 and GmPDIL-2, respectively. | ễFEBS Journal Molecular cloning and characterization of two soybean protein disulfide isomerases as molecular chaperones for seed storage proteins Shinya Kamauchi1 f Hiroyuki Wadahama1 z Kensuke Iwasaki1 Yumi Nakamoto2 Keito Nishizawa2 Masao Ishimoto2 Teruo Kawada1 and Reiko Urade1 1 Graduate Schoolof Agriculture Kyoto University Uji Japan 2 NationalAgriculturalResearch Center for Hokkaido Region Sapporo Japan Keywords endoplasmic reticulum molecular chaperone protein disulfide isomerase soybean storage protein Correspondence R. Urade Graduate Schoolof Agriculture Kyoto University Uji Kyoto 611-0011 Japan Fax 81 774 38 3758 Tel 81 774 38 3757 E-mail urade@ Present address Osaka Bioscience Institute Suita Japan These authors contributed equally to this work Database The nucleotide sequence data for the cDNA of GmPDIL-1 and GmPDIL-2 and genomic GmPDIL-1 and GmPDIL-2 are available in the DDBJ EMBL GenBank databases under accession numbers AB182628 AB185851 AB300660 and AB300661 respectively Received 26 December 2007 revised 22 February 2008 accepted 18 March 2008 doi Protein disulfide isomerase family proteins play important roles in the folding of nascent polypeptides and the formation of disulfide bonds in the endoplasmic reticulum. In this study we cloned two similar protein disulfide isomerase family genes from soybean leaf Glycine max L. Merrill. cv Jack . The cDNAs encode proteins of 525 and 551 amino acids named GmPDIL-1 and GmPDIL-2 respectively. Recombinant versions of GmP-DIL-1 and GmPDIL-2 expressed in Escherichia coli exhibited oxidative refolding activity for denatured RNaseA. Genomic sequences of both GmPDIL-1 and GmPDIL-2 were cloned and sequenced. The comparison of soybean genomic sequences with those of Arabidopsis rice and wheat showed impressive conservation of exon-intron structure across plant species. The promoter sequences of GmPDIL-1 apparently contain a cis-acting regulatory element .