RRC ID 77360
Author Tupta B, Stuehr E, Sumi MP, Sweeny EA, Smith B, Stuehr DJ, Ghosh A.
Title GAPDH is involved in the heme-maturation of myoglobin and hemoglobin.
Journal FASEB J
Abstract GAPDH, a heme chaperone, has been previously implicated in the incorporation of heme into iNOS and soluble guanylyl cyclase (sGC). Since sGC is critical for myoglobin (Mb) heme-maturation, we investigated the role of GAPDH in the maturation of this globin, as well as hemoglobins α, β, and γ. Utilizing cell culture systems, we found that overexpression of wild-type GAPDH increased, whereas GAPDH mutants H53A and K227A decreased, the heme content of Mb and Hbα and Hbβ. Overexpression of wild-type GAPDH fully recovered the heme-maturation inhibition observed with the GAPDH mutants. Partial rescue was observed by overexpression of sGCβ1 but not by overexpression of a sGCΔβ1 deletion mutant, which is unable to bind the sGCα1 subunit required to form the active sGCα1β1 complex. Wild type and mutant GAPDH was found to be associated in a complex with each of the globins and Hsp90. GAPDH at endogenous levels was found to be associated with Mb in differentiating C2C12 myoblasts, and with Hbγ or Hbα in differentiating HiDEP-1 erythroid progenitor cells. Knockdown of GAPDH in C2C12 cells suppressed Mb heme-maturation. GAPDH knockdown in K562 erythroleukemia cells suppressed Hbα and Hbγ heme-maturation as well as Hb dimerization. Globin heme incorporation was not only dependent on elevated sGCα1β1 heterodimer formation, but also influenced by iron provision and magnitude of expression of GAPDH, d-aminolevulinic acid, and FLVCR1b. Together, our data support an important role for GAPDH in the maturation of myoglobin and γ, β, and α hemoglobins.
Volume 36(2)
Pages e22099
Published 2022-2-1
DOI 10.1096/fj.202101237RR
PMID 34972240
PMC PMC9239731
MeSH Glyceraldehyde-3-Phosphate Dehydrogenases / genetics Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism* HEK293 Cells Heme / genetics Heme / metabolism* Hemoglobins / genetics Hemoglobins / metabolism* Humans K562 Cells Molecular Chaperones / genetics Molecular Chaperones / metabolism* Mutation, Missense Myoglobin / genetics Myoglobin / metabolism* Sarcoglycans / genetics Sarcoglycans / metabolism
IF 4.966
Human and Animal Cells HiDEP-1(RCB4554)