A strategy for the generation of non-aggregating mutants of Anthozoa fluorescent proteins.
Recently, we cloned several fluorescent proteins of different colors homologous to Aequorea victoria green fluorescent protein, which have great biotechnological potential as in vivo markers of gene expression. However, later investigations revealed severe drawbacks in the use of novel fluorescent proteins (FPs), in particular, the formation of tetramers (tetramerization) and high molecular weight aggregates (aggregation). In this report, we employ a mutagenic approach to resolve the problem of aggregation. The elimination of basic residues located near the N-termini of FPs results in the generation of non-aggregating versions of several FPs, specifically, drFP583 (DsRed), DsRed-Timer, ds/drFP616, zFP506, zFP538, amFP486, and asFP595.
Yanushevich
Yurii G
YG
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia.
Staroverov
Dmitry B
DB
Savitsky
Alexander P
AP
Fradkov
Arkady F
AF
Gurskaya
Nadya G
NG
Bulina
Maria E
ME
Lukyanov
Konstantin A
KA
Lukyanov
Sergey A
SA
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D003063:Cnidaria / Q000737:chemistry / Q000235:genetics; D003116:Color; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D011485:Protein Binding; D020836:Protein Structure, Quaternary
D008164:Luminescent Proteins