Tangled up in fibers: How a lytic polysaccharide monooxygenase binds its chitin substrate

Henrik Vinther Sørensen

Submitted to SASBDB: 2024 Nov 19 Published in SASBDB: 2025 Feb 9

SASDVZ9 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 66 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, 66% v/v D₂O, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDW22 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 80 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, 80% v/v D₂O, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDW32 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 100% D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, 100% v/v D₂O, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDW42 – 3 mg/ml β-chitin nanofibers from squid pens, SANS in 100 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetate, 99.7% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDW52 – 3 mg/ml β-chitin nanofibers from squid pens, SANS in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2019 Sep 21

SASDW62 – 3 mg/ml β-chitin nanofibers from squid pens in 47 % D₂O - USANS Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at BT5 USANS, NIST Center for Neutron Research on 2020 Dec 4

SASDW72 – Non-deuterated N-acetylglucosamine binding Protein A (H-GbpA) in 47 % D₂O, 2.5 mg/mL Sample: GlcNAc-binding protein A monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 100 mM NaCl, 20 mM Tris, 47% v/v D₂O, pH: 8 Experiment: SANS data collected at D11, ILL on 2020 Aug 18 RgGuinier 25.0 nm

SASDW82 – Perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O, 2.4 mg/mL Sample: GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 100 mM NaCl, 20 mM Tris, 47% v/v D₂O, pH: 8 Experiment: SANS data collected at D11, ILL on 2019 Sep 21 RgGuinier 3.7 nm Dmax 12.7 nm

SASDW92 – 10 mg/mL β-chitin nanofibers from squid pens with 2.5 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDWA2 – 3 mg/mL β-chitin nanofibers from squid pens with 2.5 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDWB2 – 3 mg/mL β-chitin nanofibers from squid pens with 2.5 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O - USANS Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at BT5 USANS, NIST Center for Neutron Research on 2020 Dec 4

SASDWC2 – 2.4 mg/mL β-chitin nanofibers from squid pens with 2 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2019 Sep 21

SASDWD2 – 2.4 mg/mL β-chitin nanofibers from squid pens with 0.5 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2019 Sep 21

SASDWE2 – 1.2 mg/mL β-chitin nanofibers from squid pens with 1 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 72 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM Acetate, 72% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2019 Sep 21

SASDWF2 – 0.48 mg/mL β-chitin nanofibers from squid pens with 1.2 mg/mL perdeuterated N-acetylglucosamine binding Protein A (D-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2019 Sep 21

SASDWG2 – 3 mg/mL β-chitin nanofibers from squid pens with 2.5 mg/mL non-deuterated N-acetylglucosamine binding Protein A (H-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDWH2 – 3 mg/mL β-chitin nanofibers from squid pens with a mixture of 1.25 mg/mL perdeuterated and 1.25 mg/mL non-deuterated N-acetylglucosamine binding Protein A (D-GbpA/H-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein GlcNAc-binding protein A monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDWJ2 – 3 mg/mL β-chitin nanofibers from squid pens with a mixture of 0.625 mg/mL perdeuterated and 1.875 mg/mL non-deuterated N-acetylglucosamine binding Protein A (D-GbpA/H-GbpA) in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid GlcNAc-binding protein A (perdeuterated) monomer, 51 kDa Vibrio cholerae serotype … protein GlcNAc-binding protein A monomer, 51 kDa Vibrio cholerae serotype … protein Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18

SASDVV9 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in H₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDVW9 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 20 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, 20% v/v D₂O, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDVX9 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 42 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetic acid, 42% v/v D₂O, pH: 3.2 Experiment: SANS data collected at D11, ILL on 2019 Jul 5

SASDVY9 – 10 mg/ml β-chitin nanofibers from squid pens, SANS in 47 % D₂O Sample: Β-chitin nanofibers from squid pens None, Squid Buffer: 20 mM acetate, 47% v/v D₂O, pH: 5 Experiment: SANS data collected at D11, ILL on 2020 Aug 18