Functional Impact of a Cancer-Related Variant in Human Δ1-Pyrroline-5-Carboxylate Reductase 1

Daudu, Oseeyi I.; Meeks, Kaylen R.; Zhang, Lu; Seravalli, Javier; Tanner, John J.; Becker, Donald F.

doi:10.1021/acsomega.2c07788

Functional Impact of a Cancer-Related Variant in Human Δ¹-Pyrroline-5-Carboxylate Reductase 1

Journal Article · Tue Jan 10 00:00:00 EST 2023 · ACS Omega

DOI:https://doi.org/10.1021/acsomega.2c07788· OSTI ID:2423441

Daudu, Oseeyi I. ^[1]; ^[2]; Zhang, Lu ^[1]; Seravalli, Javier ^[1]; ^[2]; ^[1]

Univ. of Nebraska, Lincoln, NE (United States)
Univ. of Missouri, Columbia, MO (United States)

Pyrroline-5-carboxylate reductase (PYCR) is a proline biosynthetic enzyme that catalyzes the NAD(P)H-dependent reduction of Δ¹-pyrroline-5-carboxylate (P5C) to proline. Humans have three PYCR isoforms, with PYCR1 often upregulated in different types of cancers. Here, we studied the biochemical and structural properties of the Thr171Met variant of PYCR1, which is found in patients with malignant melanoma and lung adenocarcinoma. Although PYCR1 is strongly associated with cancer progression, characterization of a PYCR1 variant in cancer patients has not yet been reported. Thr171 is conserved in all three PYCR isozymes and is located near the P5C substrate binding site. We found that the amino acid replacement does not affect thermostability but has a profound effect on PYCR1 catalytic activity. The k_cat of the PYCR1 variant T171M is 100- to 200-fold lower than wild-type PYCR1 when P5C is the variable substrate, and 10- to 25-fold lower when NAD(P)H is varied. A 1.84 Å resolution X-ray crystal structure of T171M reveals that the Met side chain invades the P5C substrate binding site, suggesting that the catalytic defect is due to steric clash preventing P5C from achieving the optimal pose for hydride transfer from NAD(P)H. These results suggest that any impact on PYCR1 function associated with T171M in cancer does not derive from increased catalytic activity.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 2423441

Journal Information:: ACS Omega, Journal Name: ACS Omega Journal Issue: 3 Vol. 8; ISSN 2470-1343

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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