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Reconstruction of the Ancestral Sequence of Calsequestrin

Reconstruction of the Ancestral Sequence of Calsequestrin. Tatyana Nienow BIOL 7020 April 26, 2010. Calsequestrin. Calcium-binding protein found in sarcoplasmic reticulum in muscle High capacity (40-80 mol Ca2+/molecule) Medium affinity (KD = 1 mM) Aspartic acid-rich tail. Calsequestrin.

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Reconstruction of the Ancestral Sequence of Calsequestrin

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  1. Reconstruction of the Ancestral Sequence of Calsequestrin Tatyana Nienow BIOL 7020 April 26, 2010

  2. Calsequestrin • Calcium-binding protein found in sarcoplasmic reticulum in muscle • High capacity (40-80 mol Ca2+/molecule) • Medium affinity (KD = 1 mM) • Aspartic acid-rich tail

  3. Calsequestrin • CSQ1 • Found in skeletal and cardiac muscle • Binds twice as many calcium ions as CSQ2 • Larger negative surface potential • CSQ2 • Found only in cardiac muscle

  4. Methods • Collected calsequestrin sequences from NCBI • Using refseq_rna database • Aligned in Seaview • Removed predicted signal sequences and poly-Asp tails

  5. Methods • Phylogeny with MrBayes and RAxML • Determined ancestral sequence with MrBayes and FastML • Corrected ancestral sequence length • Convert alignment to treat gaps as characters • Calculate ancestral form • Use ancestral gap sequence to remove inserted sites from original ancestral sequence

  6. CSQ1

  7. CSQ2

  8. Completed Ancestral Sequences

  9. Comparison to CSQ1 and CSQ2

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