scholarly journals Evaluation of Antifouling Potential and Ecotoxicity of Secondary Metabolites Derived from Red Algae of the Genus Laurencia

Marine Drugs ◽  
2019 ◽  
Vol 17 (11) ◽  
pp. 646
Author(s):  
Maria Protopapa ◽  
Manto Kotsiri ◽  
Sofoklis Mouratidis ◽  
Vassilios Roussis ◽  
Efstathia Ioannou ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Red Algae ◽  
Biological Activities ◽  
Screening Tests ◽  
Screening Process ◽  
Antifouling Activity ◽  
Mammalian Cell Lines ◽  
Vertebrate Cell

Red algae of the genus Laurencia are known to biosynthesize and secrete an immense variety of secondary metabolites possessing a spectrum of biological activities against bacteria, invertebrates and mammalian cell lines. Following a rigorous cross-species screening process, herein we report the antifouling potential of 25 secondary metabolites derived from species of the genus Laurencia, as well as the thorough evaluation of the ecotoxicity of selected metabolites against non-target marine arthropods and vertebrate cell lines. A number of these secondary metabolites exhibited potent antifouling activity and performed well in all screening tests. Our results show that perforenol (9) possesses similar antifouling activity with that already described for bromosphaerol, which is used herein as a benchmark.

scholarly journals Structure-guided selection of puromycin N-acetyltransferase mutants with enhanced selection stringency for deriving mammalian cell lines expressing recombinant proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alessandro T. Caputo ◽  
Oliver M. Eder ◽  
Hana Bereznakova ◽  
Heleen Pothuis ◽  
Albert Ardevol ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Recombinant Proteins ◽  
Cultured Cells ◽  
Hek293 Cells ◽  
Reaction Products ◽  
Enzyme Form ◽  
X Ray Crystallography ◽  
Mammalian Cell Lines ◽  
Apparent Loss

AbstractPuromycin and the Streptomyces alboniger-derived puromycin N-acetyltransferase (PAC) enzyme form a commonly used system for selecting stably transfected cultured cells. The crystal structure of PAC has been solved using X-ray crystallography, revealing it to be a member of the GCN5-related N-acetyltransferase (GNAT) family of acetyltransferases. Based on structures in complex with acetyl-CoA or the reaction products CoA and acetylated puromycin, four classes of mutations in and around the catalytic site were designed and tested for activity. Single-residue mutations were identified that displayed a range of enzymatic activities, from complete ablation to enhanced activity relative to wild-type (WT) PAC. Cell pools of stably transfected HEK293 cells derived using two PAC mutants with attenuated activity, Y30F and A142D, were found to secrete up to three-fold higher levels of a soluble, recombinant target protein than corresponding pools derived with the WT enzyme. A third mutant, Y171F, appeared to stabilise the intracellular turnover of PAC, resulting in an apparent loss of selection stringency. Our results indicate that the structure-guided manipulation of PAC function can be utilised to enhance selection stringency for the derivation of mammalian cell lines secreting elevated levels of recombinant proteins.

RAS p21 and other Gn proteins are detected in mammalian cell lines by [γ-35S]GTPγS binding

1989 ◽  
Vol 159 (3) ◽  
pp. 1269-1274 ◽  
Author(s):  
J.G. Comerford ◽  
J.R. Gibson ◽  
A.P. Dawson ◽  
I. Gibson
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines ◽  
Gtpγs Binding ◽  
Ras P21

scholarly journals Protocol to identify centrosome-associated transcription factors during mitosis in mammalian cell lines

2021 ◽  
Vol 2 (3) ◽  
pp. 100495
Author(s):  
Guobin Xie ◽  
Yuqi Zhou ◽  
Mingxuan Du ◽  
Qiqiang Wang ◽  
Jiajin Yi ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines
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