Casein Kinase 1δ-dependent Wee1 Protein Degradation

2014 ◽  
Vol 289 (27) ◽  
pp. 18893-18903 ◽  
Clara Penas ◽  
Vimal Ramachandran ◽  
Scott Simanski ◽  
Choogon Lee ◽  
Franck Madoux ◽  
1970 ◽  
Vol 24 (03/04) ◽  
pp. 325-333 ◽  
G. H Tishkoff ◽  
L. C Williams ◽  
D. M Brown

SummaryAs a corollary to our previous studies with bovine prothrombin, we have initiated a study of human prothrombin complex. This product has been isolated in crystalline form as a barium glycoprotein interaction product. Product yields were reduced compared to bovine product due to the increased solubility of the barium glycoprotein interaction product. On occasion the crystalline complex exhibited good yields. The specific activity of the crystalline complex was 1851 Iowa u/mg. Further purification of human prothrombin complex was made by removal of barium and by chromatography on Sephadex G-100 gels. The final product evidenced multiple procoagulant activities (II, VII, IX and X). The monomeric molecular weight determined by sedimentation equilibrium in a solvent of 6 M guanidine-HCl and 0.5% mercaptoethanol was 70,191 ± 3,057 and was homogeneous with respect to molecular weight. This product was characterized in regard to physical constants and chemical composition. In general, the molecular properties of human prothrombin complex are very similar to the comparable bovine product. In some preparations a reversible proteolytic enzyme inhibitor (p-aminophenylarsonic acid) was employed in the ultrafiltration step of the purification scheme to inhibit protein degradation.

2003 ◽  
Vol 89 (4) ◽  
pp. 509-515 ◽  
Patricia M. Heavey ◽  
Shirley-Anne H. Savage ◽  
Alison Parrett ◽  
Cinzia Cecchini ◽  
Christine A. Edwards ◽  

The aim of the present study was to determine the effects of age and diet (breast milk, formula milk and weaning diet) on metabolic activities in faecal samples from infants aged 1 week to 1 year, and to compare these findings with activities found in samples from adults. Such activities can provide valuable information on functional changes in the microbiota that may have significance for the health of the host. Fresh faecal samples were collected from forty-four breast-fed infants (twenty-four males, twenty females) and thirteen formula-fed infants (three males, ten females) throughout the first year of life. The samples were analysed for protein-breakdown products, including the faecal concentrations of NH3, phenol and p-cresol, and faecal bacterial enzyme activities. There was wide individual variation in all variables measured; however, the values in infants were substantially lower then those found in adults. In pre-weaned infants, faecal NH3 concentration and β-glucuronidase activity were the only endpoints that were significantly different in breast-fed and formula-fed infants (P<0·001 and P<0·05 respectively). This was not apparent after weaning. There was a significant difference between the breast-fed and formula-fed weaned groups and their pre-weaned counterparts only for NH3 (P<0·05). β-Glucuronidase activity and phenol concentration were significantly (P<0·01) greater in weaned breast-fed infants compared with pre-weaned breast-fed infants. No differences were observed between pre-weaned and weaned formula-fed infants for any of the variables except for NH3 concentration. It can be concluded from the present study that there are significant differences in two faecal characteristics between breast- and formula-fed infants and that changes occur as the infants grow older and are weaned onto solid foods.

2020 ◽  
Vol 19 (12) ◽  
pp. 1997-2014
Yadong Yu ◽  
Haichuan Liu ◽  
Zanlin Yu ◽  
H. Ewa Witkowska ◽  
Yifan Cheng

AAA+ ATPases constitute a large family of proteins that are involved in a plethora of cellular processes including DNA disassembly, protein degradation and protein complex disassembly. They typically form a hexametric ring-shaped structure with six subunits in a (pseudo) 6-fold symmetry. In a subset of AAA+ ATPases that facilitate protein unfolding and degradation, six subunits cooperate to translocate protein substrates through a central pore in the ring. The number and type of nucleotides in an AAA+ ATPase hexamer is inherently linked to the mechanism that underlies cooperation among subunits and couples ATP hydrolysis with substrate translocation. We conducted a native MS study of a monodispersed form of PAN, an archaeal proteasome AAA+ ATPase, to determine the number of nucleotides bound to each hexamer of the WT protein. We utilized ADP and its analogs (TNP-ADP and mant-ADP), and a nonhydrolyzable ATP analog (AMP-PNP) to study nucleotide site occupancy within the PAN hexamer in ADP- and ATP-binding states, respectively. Throughout all experiments we used a Walker A mutant (PANK217A) that is impaired in nucleotide binding as an internal standard to mitigate the effects of residual solvation on mass measurement accuracy and to serve as a reference protein to control for nonspecific nucleotide binding. This approach led to the unambiguous finding that a WT PAN hexamer carried – from expression host – six tightly bound ADP molecules that could be exchanged for ADP and ATP analogs. Although the Walker A mutant did not bind ADP analogs, it did bind AMP-PNP, albeit at multiple stoichiometries. We observed variable levels of hexamer dissociation and an appearance of multimeric species with the over-charged molecular ion distributions across repeated experiments. We posit that these phenomena originated during ESI process at the final stages of ESI droplet evolution.

2004 ◽  
Vol 279 (46) ◽  
pp. 48426-48433 ◽  
Zachary B. Mackey ◽  
Theresa C. O'Brien ◽  
Doron C. Greenbaum ◽  
Rebecca B. Blank ◽  
James H. McKerrow

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