Expression and functional characterization of the first bacteriophage-encoded chaperonin.
Chaperonins promote protein folding in vivo and are ubiquitously found in bacteria, archaea, and eukaryotes. The first viral chaperonin GroEL ortholog, gene product 146 (gp146), whose gene was earlier identified in the genome of bacteriophage EL, has been shown to be synthesized during phage propagation in Pseudomonas aeruginosa cells. The recombinant gp146 has been expressed in Escherichia coli and characterized by different physicochemical methods for the first time. Using serum against the recombinant protein, gp146's native substrate, the phage endolysin gp188, has been immunoprecipitated from the lysate of EL-infected bacteria and identified by mass spectrometry. In vitro experiments have shown that gp146 has a protective effect against endolysin thermal inactivation and aggregation, providing evidence of its chaperonin function. The phage chaperonin has been found to have the architecture and some properties similar to those of GroEL but not to require cochaperonin for its functional activity.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia. lpk@ibch.ru.
Lidia P
LP
Kurochkina
Pavel I
PI
Semenyuk
Victor N
VN
Orlov
Johan
J
Robben
Nina N
NN
Sykilinda
Vadim V
VV
Mesyanzhinov
D001483:Base Sequence; D018833:Chaperonins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004279:DNA, Viral / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D046912:Multiprotein Complexes; D011489:Protein Denaturation; D055503:Protein Multimerization; D017105:Pseudomonas Phages / Q000235:genetics* / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000821:virology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D004279:DNA, Viral; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; D014764:Viral Proteins; D018833:Chaperonins