Journal Article

NADP-malic Enzyme and Hsp70: Co-purification of Both Proteins and Modification of NADP-malic Enzyme Properties by Association with Hsp70

María V. Lara, María F. Drincovich, Gabriela L. Müller, Verónica G. Maurino and Carlos S. Andreo

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 6, pages 997-1006
Published in print June 2005 | ISSN: 0032-0781
Published online June 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci108
NADP-malic Enzyme and Hsp70: Co-purification of Both Proteins and Modification of NADP-malic Enzyme Properties by Association with Hsp70

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Different preparations of antibodies against 62 kDa NADP-malic enzyme (NADP-ME) from purified maize leaves cross-react with a 72 kDa protein from diverse tissues in many species. A 72 kDa protein, suggested to be a non-photosynthetic NADP-ME, has been purified from several plant species. However, to date, a cDNA coding for this putative 72 kDa NADP-ME has not been isolated. The screening of maize and tobacco leaf expression libraries using antibodies against purified 62 kDa NADP-ME allowed the identification of a heat shock protein (Hsp70). In addition, tandem mass spectrometry (MS/MS) studies indicate that along with NADP-ME, a 72 kDa protein, identified as an Hsp70 and reacting with the antibodies, is also purified from maize roots. On the other hand, the screening of a maize root cDNA library revealed the existence of a cDNA that encodes a mature 66 kDa NADP-ME. These results suggest that the 72 kDa protein is not actually an NADP-ME but in fact an Hsp70, at least in maize and tobacco. Probably, NADP-ME–Hsp70 association, taking place at least when preparing crude extracts, can lead to a co-purification of the proteins and can thus explain the cross-reaction of the antibodies. In the present work, we analyse and discuss a probable interaction of NADP-ME with Hsp70.

Keywords: Hsp70; NADP-malic enzyme; Nicotiana tabacum; Protein interaction; Zea mays; a-p62 NADP-ME, antibodies against purified 62 kDa NADP-ME from maize leaves; a-r62 NADP-ME, antibodies against recombinant 62 kDa NADP-ME; a-r66 NADP-ME, antibodies against recombinant 66 kDa NADP-ME; a-Hsp70T, antibodies against recombinant Hsp70 from tobacco leaves; BSA, bovine seum albumin; GFP, green fluorescent protein; Hsp70, heat shock protein 70; MS, mass spectrometry; NADP-ME, NADP-malic enzyme; Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygenase

Journal Article.  7371 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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