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Plant and Cell Physiology Advance Access published online on April 19, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci108
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received February 9, 2005
Accepted April 9, 2005

Regular Paper

NADP-Malic Enzyme and Hsp70: Co-Purification of Both Proteins and Modification of NADP-Malic Enzyme Properties by Association with Hsp70

María V. Lara 1, María F. Drincovich 1, Gabriela L. Müller 1, Verónica G. Maurino 2, and Carlos S. Andreo 1*

1 Centro de Estudios Fotosintéticos y Bioquímicos, Facultad de Ciencias Bioquímicas y Farmacéuticas, Suipacha 531, Rosario (2000), Argentina
2 Botanisches Institut, Universität zu Köln, Gyrhofstr, 15, D-50931, Cologne, Germany

* To whom correspondence should be addressed.
Carlos S. Andreo, E-mail: candreo{at}fbioyf.unr.edu.ar


  Abstract

Different preparations of antibodies against purified maize leaves 62 kDa NADP-malic enzyme (NADP-ME) 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, until present a cDNA encoding 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, 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 (Saigo et al. 2004). These results suggest that the 72 kDa protein is not actually a 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, 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.
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