The effects of pyrroloquinoline quinone on heme-regulated eIF-2alpha kinase and eIF-2B activities in eukaryotic protein synthesis.
Pyrroloquinoline quinone (PQQ), a novel cofactor of biological redox processes, is ubiquitous in animal cells. We have examined the effects of PQQ on protein synthesis. PQQ inhibits protein synthesis in hemin-supplemented rabbit reticulocyte lysates. This inhibition is characterized by increased phosphorylation of eIF-2alpha and by diminished guanine nucleotide exchange activity of eIF-2B. The increased eIF-2alpha phosphorylation is the result of activation by PQQ of the heme-regulated eIF-2alpha kinase (HRI). The addition of 10 microM PQQ completely inhibits the increase in protein synthesis that occurs on the addition of hemin (20 microM) to heme-deficient lysates, whereas a lower concentration of PQQ (100 nM) causes a very slight stimulation of protein synthesis. The increased eIF-2alpha phosphorylation that occurs at high concentrations of PQQ inhibits eIF-2B activity, presumably due to formation of a 15S complex [eIF-2(alphaP).eIF-2B] in which eIF-2B becomes non-functional. Low concentrations of PQQ (0.1-1 microM) do not affect eIF-2alpha phosphorylation, but rather enhance the guanine nucleotide exchange activity of eIF-2B in reticulocyte lysates. In Chinese hamster ovary cell extract which is devoid of significant eIF-2alpha kinase activity, addition of both low and high concentrations of PQQ results in an increase in eIF-2B activity. The addition of PQQ to reticulocyte lysates activates HRI whereas addition of PQQ to purified HRI in vitro inhibits the autokinase and eIF-2alpha kinase activity of the HRI; the inhibition of purified HRI by PQQ is observed both in the presence and absence of hemin. These findings suggest that PQQ inhibits purified HRI by acting as an oxidant whereas in lysates in which PQQ is readily reduced, the PQQ acts as a reductant and increases the activities of both HRI and eIF-2B.
Atchuta Ramaiah
K V
KV
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, E25-551, Cambridge, MA 02139, USA.
Chen
J J
JJ
Gallop
P M
PM
London
I M
IM
D000818:Animals; D016466:CHO Cells; D002474:Cell-Free System; D003067:Coenzymes / Q000494:pharmacology; D006224:Cricetinae; D004789:Enzyme Activation; D020662:Guanine Nucleotide Exchange Factors; D006153:Guanosine Diphosphate / Q000378:metabolism; D006427:Hemin / Q000494:pharmacology*; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D045542:PQQ Cofactor; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism*; D015363:Quinolones / Q000494:pharmacology*; D011809:Quinones / Q000494:pharmacology*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D019892:eIF-2 Kinase / Q000096:biosynthesis*
D003067:Coenzymes; D020662:Guanine Nucleotide Exchange Factors; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D015363:Quinolones; D011809:Quinones; D006153:Guanosine Diphosphate; D045542:PQQ Cofactor; D006427:Hemin; D019892:eIF-2 Kinase; D007930:Leucine