Massively parallel, computationally guided design of a proenzyme.

Yachnin BJ, Azouz LR, White RE 3rd, Minetti CASA, Remeta DP, Tan VM, Drake JM, Khare SD
Proc Natl Acad Sci U S A 119(15):e2116097119 (2022 Apr 12)

PMID: 35377786 doi: 10.1073/pnas.2116097119

Submitted to SASBDB: 2021 Mar 23 Published in SASBDB: 2021 Apr 7

SASDLD2 – Circular permutant Carboxypeptidase G2-CP-N89-K177A Sample: Carboxypeptidase G2 (circular permutant CP-N89) K177A dimer, 85 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2017 Jun 6 RgGuinier 3.6 nm Dmax 14.0 nm VolumePorod 105 nm3

SASDLE2 – Circular permutant Carboxypeptidase G2-CP-N89-K177A with methotrexate Sample: Carboxypeptidase G2 (circular permutant CP-N89) K177A dimer, 85 kDa Pseudomonas sp. (strain … protein Methotrexate dimer, 1 kDa Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2017 Jun 6 RgGuinier 3.5 nm Dmax 12.7 nm VolumePorod 104 nm3

SASDLF2 – Pro-CPG2-1 (pro-enzyme design 1 of circular permutant Carboxypeptidase G2-CP-N89-K177A) Sample: Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 1 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.6 nm Dmax 12.4 nm VolumePorod 125 nm3

SASDLG2 – Pro-CPG2-1 (pro-enzyme design 1 of circular permutant Carboxypeptidase G2-CP-N89-K177A) with Methotrexate Sample: Methotrexate dimer, 1 kDa Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 1 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.5 nm Dmax 12.2 nm VolumePorod 123 nm3

SASDLH2 – Pro-CPG2-2 (pro-enzyme design 2 of circular permutant Carboxypeptidase G2-CP-N89-K177A) Sample: Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 2 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.7 nm Dmax 12.5 nm VolumePorod 126 nm3

SASDLJ2 – Pro-CPG2-2 (pro-enzyme design 2 of circular permutant Carboxypeptidase G2-CP-N89-K177A) with Methotrexate Sample: Methotrexate dimer, 1 kDa Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 2 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.5 nm Dmax 12.1 nm VolumePorod 125 nm3

SASDLK2 – Pro-CPG2-3 (pro-enzyme design 3 of circular permutant Carboxypeptidase G2-CP-N89-K177A) Sample: Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 3 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.7 nm Dmax 13.0 nm VolumePorod 127 nm3

SASDLL2 – Pro-CPG2-3 (pro-enzyme design 3 of circular permutant Carboxypeptidase G2-CP-N89-K177A) with Methotrexate Sample: Methotrexate dimer, 1 kDa Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 3 dimer, 100 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 May 8 RgGuinier 3.5 nm Dmax 12.3 nm VolumePorod 125 nm3

SASDLM2 – Pro-CPG2-1-Disulfide (pro-enzyme design 1 disulfide variant of circular permutant Carboxypeptidase G2-CP-N89-K177A) Sample: Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 1 Disulfide Variant dimer, 96 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 Dec 6 RgGuinier 3.6 nm Dmax 12.6 nm VolumePorod 120 nm3

SASDLN2 – Pro-CPG2-1-Disulfide (pro-enzyme design 1 disulfide variant of circular permutant Carboxypeptidase G2-CP-N89-K177A) with Methotrexate Sample: Methotrexate dimer, 1 kDa Pro-Carboxypeptidase G2 (circular permutant CP-N89) K177A Design 1 Disulfide Variant dimer, 96 kDa Pseudomonas sp. (strain … protein Buffer: 50 mM Tris, 100 mM NaCl, pH: 7.4 Experiment: SAXS data collected at 12.3.1 (SIBYLS), Advanced Light Source (ALS) on 2018 Dec 6 RgGuinier 3.5 nm Dmax 12.3 nm VolumePorod 125 nm3