Expression level of Ubc9 protein in rat tissues.

• Authors: Filip Gołębiowski , Aneta Szulc , Monika Sakowicz , Andrzej Szutowicz , Tadeusz Pawełczyk
• Languages of publication: EN

Abstracts

EN

Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDa band of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein appears to have some degree of tissue specificity.

Keywords

EN

Ubc9; rat; recombinant protein; tissue distribution

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2003
• Volume: 50
• Issue: 4
• Pages: 1064-1073

Dates

published

2003

received

2003-09-27

revised

2003-11-28

accepted

2003-12-09

Contributors

author

Filip Gołębiowski

• Department of Molecular Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland

author

Aneta Szulc

• Department of Molecular Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland

author

Monika Sakowicz

• Department of Molecular Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland

author

Andrzej Szutowicz

• Department of Laboratory Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland

author

Tadeusz Pawełczyk

• Department of Molecular Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland

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