Abstract
Bacterial ribosomal L7/L12 stalk is formed by L10, L11, and multiple copies of L7/L12, which plays an essential role in recruiting initiation and elongation factors during translation. The homologs of these proteins, MRPL10, MRPL11, and MRPL12, are present in human mitochondrial ribosomes. To evaluate the role of MRPL10, MRPL11, and MRPL12 in translation, we over-expressed and purified components of the human mitochondrial L7/L12 stalk proteins in Escherichia coli. Here, we designed a construct to co-express MRPL10 and MRPL12 using a duet expression system to form a functional MRPL10-MRPL12 complex. The goal is to demonstrate the homology between the mitochondrial and bacterial L7/L12 stalk proteins and to reconstitute a hybrid ribosome to be used in structural and functional studies of the mitochondrial stalk.
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Animals
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Blotting, Western
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Cattle
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Cell Cycle Proteins / chemistry*
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / isolation & purification
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Cell Cycle Proteins / metabolism
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Chromatography, Affinity
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Escherichia coli / chemistry*
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Escherichia coli Proteins / chemistry*
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Escherichia coli Proteins / metabolism
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Humans
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Molecular Sequence Data
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Nuclear Proteins / chemistry*
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Nuclear Proteins / genetics
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Nuclear Proteins / isolation & purification
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Nuclear Proteins / metabolism
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Potassium Chloride / chemistry
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Ribosomal Proteins / chemistry*
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Ribosomal Proteins / genetics
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Ribosomal Proteins / isolation & purification
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Ribosomal Proteins / metabolism
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Ribosomes / chemistry*
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Ribosomes / metabolism
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Sequence Alignment
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Solubility
Substances
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Cell Cycle Proteins
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Escherichia coli Proteins
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MRPL12 protein, human
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Nuclear Proteins
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Ribosomal Proteins
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Potassium Chloride