Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Palmitoylation of the pore-forming subunit of Ca(v)1.2 controls channel voltage sensitivity and calcium transients in cardiac myocytes

doi: 10.1073/pnas.2207887120

Figure Lengend Snippet: Ca(v) 1.2 α1C subunit is palmitoylated in mouse, rabbit, and human ventricular tissues. Palmitoylated proteins were purified by resin-assisted capture of acylated proteins (acyl-RAC) and immunoblotted as shown. The bar chart below each blot indicates the abundance of Ca(v)1.2 α1C and caveolin 3 (Cav3) in the purified palmitoylated fraction (Palm) relative to the corresponding unfractionated lysate (UF). N = 4 (mouse), N = 8 (rabbit), N = 7 (human). *** P < 0.001, unpaired t test. Error bars represent SEM.

Article Snippet: Anti-Ca(v)1.2 α1C subunit antibodies raised in rabbit and guinea pig were obtained from Alomone Labs, antibodies against flotillin 2 and caveolin 3 from BD Biosciences, and anti-GFP antibodies from Abcam and Protein Tech.

Techniques: Purification

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Palmitoylation of the pore-forming subunit of Ca(v)1.2 controls channel voltage sensitivity and calcium transients in cardiac myocytes

doi: 10.1073/pnas.2207887120

Figure Lengend Snippet: Palmitoylation site conservation in human Ca(v)1 channel isoforms. For clarity, regions of the rabbit α1C splice variant of Ca(v)1.2 CACH2A (UniProt accession number P15381) with palmitoylated cysteines identified in this investigation numbered and highlighted in red are shown above the corresponding regions of the human channels. Numbers at the end of each sequence are the numbering of the final amino acid in the region of each Ca(v)1 isoform shown. Human Ca(v)1 isoforms (UniProt accession numbers shown) were aligned using Clustal Ω. “*” below an amino acid indicates 100% conservation between isoforms; “:” indicates amino acids of highly similar properties; “.” indicates amino acids of weakly similar properties. The palmitoylation site in the Ca(v)1.2 N terminus is conserved in all isoforms. Ca(v)1.1 does not possess a cysteine analogous to C519 in the I–II linker, but Ca(v)1.3 and 1.4 do. C543 is unique to Ca(v)1.2.

Article Snippet: Anti-Ca(v)1.2 α1C subunit antibodies raised in rabbit and guinea pig were obtained from Alomone Labs, antibodies against flotillin 2 and caveolin 3 from BD Biosciences, and anti-GFP antibodies from Abcam and Protein Tech.

Techniques: Variant Assay, Sequencing

Journal: Molecular Neurobiology

Article Title: Involvement of the Voltage-Gated Calcium Channels L- P/Q- and N-Types in Synapse Elimination During Neuromuscular Junction Development

doi: 10.1007/s12035-022-02818-2

Figure Lengend Snippet: a Representative confocal image of a nerve terminal arborization. Singly, dually, and innervated by three or more axons NMJs from YFP muscles and also images of the morphologic maturation (S1, the most inmature, and S4, almost fully differentiated, stages) of the postsynaptic clusters from P9 mice. The bar indicates 10 μm. b Confocal immunofluorescence location of α 1D L-, N-, and P/Q-type voltage-dependent calcium channels (VDCCs) at the NMJ. Triple labeling of VDCCs (green fluorescence) with syntaxin (blue fluorescence) and nAChR-α-bungarotoxin (red fluorescence) in merge images. Figure shows the presence of α 1D L-, N-, and P/Q-type-VDCC (in green) in the nerve terminal of P9 Levator auris longus (LAL) muscle endplates. The bar indicates 10 μm

Article Snippet: Muscles were incubated overnight at 4 °C with anti-Ca V 1.3 (CACNA1D) antibody voltage-dependent L-type calcium channel subunit α 1D (1/100; ACC-005, Alomone Labs, Jerusalem, Israel); anti-Ca V 2.1 (CACNA1A) antibody voltage-dependent P/Q-type calcium channel subunit α 1A (1/100; ACC-001, Alomone Labs, Jerusalem, Israel); anti-Ca V 2.2 (CACNA1B) antibody voltage-dependent N-type calcium channel subunit α 1B (1/100; ACC1-002, Alomone Labs, Jerusalem, Israel), and anti-mouse syntaxin (1/1000, S066, Sigma, St Louis, MO, USA).

Techniques: Immunofluorescence, Labeling, Fluorescence