Protocol detail

Structural Characterization of PKCα Domains

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The DNA sequences encoding C1B-C2 (residues 100–293), isolated C1B (residues 100–152) or C2 (residues 155–293) of PKCα (M. musculus for C1B-C2 and C1B; R. Norvegicus for C2) were amplified by PCR using the cDNA clone of PKCα (Open Biosystems) as a template and cloned into the pET-SUMO vector (Invitrogen). Isolated C1B, C2, and C1B-C2 were expressed and purified as described previously (Morales et al., 2011 (link); Stewart et al., 2011 (link); Cole et al., 2019 (link)). [U-¹⁵N, 75%-²H]-enriched C1B-C2 and [U-¹⁵N]- or [U-¹³C, ¹⁵N]-enriched C1B were used for the NMR experiments.

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Cole T.R, & Igumenova T.I. (2021). Reactivity of Thiol-Rich Zn Sites in Diacylglycerol-Sensing PKC C1 Domain Probed by NMR Spectroscopy. Frontiers in Molecular Biosciences, 8, 728711.

Publication 2021

pET-SUMO vector

Cdna Dna sequences Musculus Pkcα Vector

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Variable analysis

independent variables

The DNA sequences encoding C1B-C2 (residues 100–293), isolated C1B (residues 100–152) or C2 (residues 155–293) of PKCα

dependent variables

Isolated C1B, C2, and C1B-C2 were expressed and purified

control variables

The cDNA clone of PKCα (Open Biosystems) was used as a template
The pET-SUMO vector (Invitrogen) was used for cloning

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