PMID:15840644
| Citation |
Lara, MV, Drincovich, MF, Müller, GL, Maurino, VG and Andreo, CS (2005) NADP-malic enzyme and Hsp70: co-purification of both proteins and modification of NADP-malic enzyme properties by association with Hsp70. Plant Cell Physiol. 46:997-1006 |
|---|---|
| Abstract |
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. |
| Links |
PubMed Online version:10.1093/pcp/pci108 |
| Keywords |
Amino Acid Sequence; Base Sequence; Cross Reactions; DNA, Complementary/genetics; DNA, Plant/genetics; Enzyme Stability; Gene Library; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; HSP70 Heat-Shock Proteins/genetics; HSP70 Heat-Shock Proteins/immunology; HSP70 Heat-Shock Proteins/isolation & purification; HSP70 Heat-Shock Proteins/metabolism; Kinetics; Malate Dehydrogenase/genetics; Malate Dehydrogenase/immunology; Malate Dehydrogenase/isolation & purification; Malate Dehydrogenase/metabolism; Molecular Sequence Data; Plant Proteins/genetics; Plant Proteins/immunology; Plant Proteins/isolation & purification; Plant Proteins/metabolism; Plants, Genetically Modified; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/isolation & purification; Recombinant Fusion Proteins/metabolism; Tobacco/genetics; Tobacco/metabolism; Zea mays/genetics; Zea mays/metabolism |
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