Ligand‐induced CaMKIIα hub Trp403 flip, hub domain stacking, and modulation of kinase activity

Narayanan D, Larsen A, Gauger S, Adafia R, Hammershøi R, Hamborg L, Bruus‐Jensen J, Griem‐Krey N, Gee C, Frølund B, Stratton M, Kuriyan J, Kastrup J, Langkilde A, Wellendorph P, Solbak S
Protein Science 33(10) (2024 Sep 14)

doi: 10.1002/pro.5152

Submitted to SASBDB: 2024 Mar 18 Published in SASBDB: 2024 Sep 18

SASDUQ4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2022 Oct 3 RgGuinier 4.5 nm Dmax 15.2 nm VolumePorod 367 nm3

SASDUR4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2020 Nov 19 RgGuinier 4.4 nm Dmax 14.9 nm VolumePorod 351 nm3

SASDUS4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with PIPA (i) Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2022 Nov 30 RgGuinier 4.8 nm Dmax 16.0 nm VolumePorod 396 nm3

SASDUT4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with PIPA (ii) Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2023 May 30 RgGuinier 5.5 nm Dmax 27.0 nm

SASDUU4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with PIPA (ii-remeasured after storage) Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2023 May 31 RgGuinier 8.8 nm Dmax 37.0 nm

SASDUV4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with PIPA (iii) Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2023 May 30 RgGuinier 4.5 nm Dmax 17.0 nm VolumePorod 556 nm3

SASDUW4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with PIPA (iv) Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2023 Mar 17 RgGuinier 15.6 nm Dmax 90.0 nm

SASDUX4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with 5-HDC Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 5-hydroxydiclofenac monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 100 µM 5-HDC, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2022 Oct 3 RgGuinier 4.5 nm Dmax 17.5 nm VolumePorod 372 nm3

SASDUY4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) WT hub domain with HOCPCA Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha dodecamer, 186 kDa Homo sapiens protein 3-hydroxycyclopent-1-enecarboxylic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 1 mM HOCPCA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2020 Nov 19 RgGuinier 4.1 nm Dmax 12.7 nm VolumePorod 310 nm3

SASDUZ4 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) W403L hub domain Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) dodecamer, 185 kDa Homo sapiens protein Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2023 Mar 17 RgGuinier 4.7 nm Dmax 16.5 nm VolumePorod 358 nm3

SASDU25 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) W403L hub domain with PIPA Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) dodecamer, 185 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2023 Mar 17 RgGuinier 4.4 nm Dmax 15.0 nm VolumePorod 334 nm3

SASDU35 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) W403L hub domain with 5-HDC Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (W403L) dodecamer, 185 kDa Homo sapiens protein 5-hydroxydiclofenac monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 100 µM 5-HDC, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L, Département de Biochimie, Université de Montréal on 2022 Dec 15 RgGuinier 4.4 nm Dmax 15.0 nm VolumePorod 335 nm3

SASDU45 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) 14-mer, 217 kDa Homo sapiens protein Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, pH: 6 Experiment: SAXS data collected at EMBL P12, PETRA III on 2022 Oct 3 RgGuinier 4.7 nm Dmax 18.0 nm VolumePorod 329 nm3

SASDU55 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) 14-mer, 217 kDa Homo sapiens protein Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2020 Nov 19 RgGuinier 4.5 nm Dmax 14.5 nm VolumePorod 313 nm3

SASDU65 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain with PIPA Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) 14-mer, 217 kDa Homo sapiens protein 2-(6-(4-chlorophenyl)imidazo[1,2-b]pyridazine-2-yl)acetic acid monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 200 µM PIPA, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2022 Nov 30 RgGuinier 4.7 nm Dmax 20.0 nm VolumePorod 375 nm3

SASDU75 – Calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) 6x hub domain with 5-HDC Sample: Calcium/calmodulin-dependent protein kinase type II subunit alpha (T354N, E355Q, T412N, I414M, I464H, F467M) 14-mer, 217 kDa Homo sapiens protein 5-hydroxydiclofenac monomer, 0 kDa Buffer: 20 mM MES, 150 mM NaCl, 1 mM DTT, 100 µM 5-HDC, pH: 6 Experiment: SAXS data collected at Xenocs BioXolver L with MetalJet, University of Copenhagen, Department of Drug Design and Pharmacology on 2020 Nov 19 RgGuinier 4.7 nm Dmax 18.5 nm VolumePorod 366 nm3