Chimeric Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase/oxygenase containing a pea small subunit protein is compromised in carbamylation
| Autor: | T P, Getzoff, G, Zhu, H J, Bohnert, R G, Jensen |
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| Rok vydání: | 1998 |
| Předmět: |
DNA
Complementary Sequence Homology Amino Acid Recombinant Fusion Proteins Ribulose-Bisphosphate Carboxylase Molecular Sequence Data Arabidopsis food and beverages Genes Plant Plants Genetically Modified Electrophoresis Gel Two-Dimensional Amino Acid Sequence Carbamates Plant Proteins Research Article |
| Zdroj: | Plant physiology. 116(2) |
| ISSN: | 0032-0889 |
| Popis: | A cDNA of pea (Pisum sativum L.) RbcS 3A, encoding a small subunit protein (S) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), has been expressed in Arabidopsis thaliana under control of the cauliflower mosaic virus 35S promoter, and the transcript and mature S protein were detected. Specific antibodies revealed two protein spots for the four Arabidopsis S and one additional spot for pea S. Pea S in chimeric Rubisco amounted to 15 to 18% of all S, as judged by separation on two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels from partially purified enzyme preparations and quantitation of silver-stained protein spots. The chimeric enzyme had 11 ± 1% fewer carbamylated sites and a 11 ± 1% lower carboxylase activity than wild-type Arabidopsis Rubisco. Whereas pea S expression, preprotein transport, and processing and assembly resulted in a stable holoenzyme, the chimeric enzyme was reproducibly catalytically less efficient. We suggest that the presence of, on average, one foreign S per holoenzyme is responsible for the altered activity. In addition, higher-plant Rubisco, unlike the cyanobacterial enzyme, seems to have evolved species-specific interactions between S and the large subunit protein that are involved in carbamylation of the active site. |
| Databáze: | OpenAIRE |
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