Expression level of Ubc9 protein in rat tissues.
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.
Gołebiowski
Filip
F
Department of Molecular Medicine, Medical University of Gdańsk, 80-211 Gdańsk, Poland.
Szulc
Aneta
A
Sakowicz
Monika
M
Szutowicz
Andrzej
A
Pawełczyk
Tadeusz
T
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D004587:Electrophoresis, Agar Gel; D006801:Humans; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013696:Temperature; D013997:Time Factors; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D012333:RNA, Messenger; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9