Long-range structural defects by pathogenic mutations in most severe glucose-6-phosphate dehydrogenase deficiency

Horikoshi, Naoki; Hwang, Sunhee; Gati, Cornelius; Matsui, Tsutomu; Castillo-Orellana, Carlos; Raub, Andrew G.; Garcia, Adriana A.; Jabbarpour, Fatemeh; Batyuk, Alexander; Broweleit, Joshua; Xiang, Xinyu; Chiang, Andrew; Broweleit, Rachel; Vöhringer-Martinez, Esteban; Mochly-Rosen, Daria; Wakatsuki, Soichi

doi:10.1073/pnas.2022790118

Long-range structural defects by pathogenic mutations in most severe glucose-6-phosphate dehydrogenase deficiency

Journal Article · Tue Jan 19 00:00:00 EST 2021 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2022790118· OSTI ID:1766136

^[1]; Hwang, Sunhee ^[2]; Gati, Cornelius ^[3]; Matsui, Tsutomu ^[4]; ^[5]; Raub, Andrew G. ^[2]; Garcia, Adriana A. ^[2]; Jabbarpour, Fatemeh ^[3]; ^[4]; Broweleit, Joshua ^[2]; ^[2]; ^[2]; Broweleit, Rachel ^[2]; Vöhringer-Martinez, Esteban ^[5]; ^[2]; ^[3]

Univ. of Tsukuba (Japan); SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., CA (United States)
Stanford Univ., CA (United States)
SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., CA (United States)
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Univ. de Concepcion (Chile)

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common blood disorder, presenting multiple symptoms, including hemolytic anemia. It affects 400 million people worldwide, with more than 160 single mutations reported in G6PD. The most severe mutations (about 70) are classified as class I, leading to more than 90% loss of activity of the wild-type G6PD. The crystal structure of G6PD reveals these mutations are located away from the active site, concentrating around the noncatalytic NADP+-binding site and the dimer interface. However, the molecular mechanisms of class I mutant dysfunction have remained elusive, hindering the development of efficient therapies. To resolve this, we performed integral structural characterization of five G6PD mutants, including four class I mutants, associated with the noncatalytic NADP+ and dimerization, using crystallography, small-angle X-ray scattering (SAXS), cryogenic electron microscopy (cryo-EM), and biophysical analyses. Comparisons with the structure and properties of the wild-type enzyme, together with molecular dynamics simulations, bring forward a universal mechanism for this severe G6PD deficiency due to the class I mutations. We highlight the role of the noncatalytic NADP+-binding site that is crucial for stabilization and ordering two β-strands in the dimer interface, which together communicate these distant structural aberrations to the active site through a network of additional interactions. This understanding elucidates potential paths for drug development targeting G6PD deficiency.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: Fondecyt; Japan Society for the Promotion of Science (KAKENHI); Max Planck Institute; National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1766136

Alternate ID(s):: OSTI ID: 1778263

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 4 Vol. 118; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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