Recombinant human Tif1γ (Trim33)

 

Catalog # : EPX-006-RBV

 

Source : Human

 

Expressed in : SF9 cells

 

Quantity : 10g of enzymatically active recombinant Tif1γ at 0.5g /l

 

 

 

Background:

 

Tif1γ (Trim33) is an E3 ubiquitin-protein ligase, which promotes SMAD4 ubiquitination, nuclear exclusion and degradation via the ubiquitin proteasome pathway (1, 2). May act as a transcriptional repressor. Inhibits the transcriptional response to TGF-beta/BMP signaling cascade. Plays a role in the control of cell proliferation. Its association with SMAD2 and SMAD3 stimulates erythroid differentiation of hematopoietic stem/progenitor By similarity. Monoubiquitinates SMAD4 and acts as an inhibitor of SMAD4-dependent TGF-beta/BMP signaling cascade (Monoubiquitination of SMAD4 hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-beta/BMP signaling cascade) (2). Defects in TRIM33 are a cause of thyroid papillary carcinoma (3).

 

 

Protein details:

 

Recombinant human N-terminal FLAG tagged Tif1γ was produced in SF9 insect cells, purified using FPLC and formulated in a storage buffer containing 20mM Tris-Cl pH 7.6, 1mM EDTA, 0.15 M NaCl, 10% glycerol, 0.5mM PMSF and 1mM DTT. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

 

 

Quality control:

 

Each lot has been evaluated by 10% Tris Glycine SDS-PAGE

 

center  

SDS-PAGE gel of recombinant FLAG tagged Tif1α (Lane 2).  Lane 1, protein molecular weight marker.



SDS-PAGE gel of recombinant auto-ubiquitylated Tif1γ (Lane 2).  Lane 1, recombinant FLAG Tagged Tif1γ.

 

Ubiquitylation assay:

 

Tif1γ modification with ubiquitin (Ub) was reconstituted in vitro in the presence of recombinant E1, E2, Ub and ATP. Tif1γ auto-ubquitylation results in a mobility shift of 8kDa. The efficiency of in vitro auto-ubiquitylation is evaluated by 8% Tris Glycine SDS-PAGE.

 

 

Storage:

 

-80C

 

 

Guarantee:

 

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

 

 

 

Sequence:


MAENKGGGEAESGGGGSGSAPVTAGAAGPAAQEAEPPLTAVLVEEEEEEGGRAGAEGGAA
GPDDGGVAAASSGSAQAASSPAASVGTGVAGGAVSTPAPAPASAPAPGPSAGPPPGPPAS
LLDTCAVCQQSLQSRREAEPKLLPCLHSFCLRCLPEPERQLSVPIPGGSNGDIQQVGVIR
CPVCRQECRQIDLVDNYFVKDTSEAPSSSDEKSEQVCTSCEDNASAVGFCVECGEWLCKT
CIEAHQRVKFTKDHLIRKKEDVSESVGASGQRPVFCPVHKQEQLKLFCETCDRLTCRDCQ
LLEHKEHRYQFLEEAFQNQKGAIENLLAKLLEKKNYVHFAATQVQNRIKEVNETNKRVEQ
EIKVAIFTLINEINKKGKSLLQQLENVTKERQMKLLQQQNDITGLSRQVKHVMNFTNWAI
ASGSSTALLYSKRLITFQLRHILKARCDPVPAANGAIRFHCDPTFWAKNVVNLGNLVIES
KPAPGYTPNVVVGQVPPGTNHISKTPGQINLAQLRLQHMQQQVYAQKHQQLQQMRMQQPP
APVPTTTTTTQQHPRQAAPQMLQQQPPRLISVQTMQRGNMNCGAFQAHQMRLAQNAARIP
GIPRHSGPQYSMMQPHLQRQHSNPGHAGPFPVVSVHNTTINPTSPTTATMANANRGPTSP
SVTAIELIPSVTNPENLPSLPDIPPIQLEDAGSSSLDNLLSRYISGSHLPPQPTSTMNPS
PGPSALSPGSSGLSNSHTPVRPPSTSSTGSRGSCGSSGRTAEKTSLSFKSDQVKVKQEPG
TEDEICSFSGGVKQEKTEDGRRSACMLSSPESSLTPPLSTNLHLESELDALASLENHVKI
EPADMNESCKQSGLSSLVNGKSPIRSLMHRSARIGGDGNNKDDDPNEDWCAVCQNGGDLL
CCEKCPKVFHLTCHVPTLLSFPSGDWICTFCRDIGKPEVEYDCDNLQHSKKGKTAQGLSP
VDQRKCERLLLYLYCHELSIEFQEPVPASIPNYYKIIKKPMDLSTVKKKLQKKHSQHYQI
PDDFVADVRLIFKNCERFNEMMKVVQVYADTQEINLKADSEVAQAGKAVALYFEDKLTEI
YSDRTFAPLPEFEQEEDDGEVTEDSDEDFIQPRRKRLKSDERPVHIK



 

References:

1.    Venturini et al., Oncogene 18:1209-1217(1999)
2.    He et al., Cell 125:929-941(2006)
3.    Greenman et al., Nature 446:153-158(2007)


 

 

 


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