Conformational stability of ribosomal protein L7/L12: effects of pH, temperature, and guanidinium chloride

Luer, Carl A.; Wong, K.P.

doi:10.1021/bi00542a027

Cited by 22 publications

(9 citation statements)

References 48 publications

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“…The L7/ L12 ribosomal protein exists as a dimer in their biologically functional form, the monomers of which are identical except for the acetylated N-terminal serine of L7. The C-terminal fragment used in this study (termed Ctf) is, however, known to be remarkably stable in the isolated form as well (37) and thus suitable as a model protein.…”

Section: Methodsmentioning

confidence: 99%

Protein Structures under Electrospray Conditions

2007

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During electrospray ionization (ESI), proteins are transferred from solution into vacuum, a process that influences the conformation of the protein. Exactly how much the conformation changes due to the dehydration process, and in what way, is difficult to determine experimentally. The aim of this study is therefore to monitor what happens to protein structures as the surrounding waters gradually evaporate, using computer simulations of the transition of proteins from water to vacuum. Five different proteins have been simulated with water shells of varying thickness, enabling us to mimic the entire dehydration process. We find that all protein structures are affected, at least to some extent, by the transfer but that the major features are preserved. A water shell with a thickness of roughly two molecules is enough to emulate bulk water and to largely maintain the solution phase structure. The conformations obtained in vacuum are quite similar and make up an ensemble which differs from the structure obtained by experimental means, and from the solution phase structure as found in simulations. Dehydration forces the protein to make more intramolecular hydrogen bonds, at the expense of exposing more hydrophobic area (to vacuum). Native hydrogen bonds usually persist in vacuum, yielding an easy route to refolding upon rehydration. The findings presented here are promising for future bio-imaging experiments with X-ray free electron lasers, and they strongly support the validity of mass spectrometry experiments for studies of intra- and intermolecular interactions.

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Section: Methodsmentioning

confidence: 99%

Protein Structures under Electrospray Conditions

2007

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“…The relative positions of the subunits in the dimer are unknown. While Gudkov et al (14) concluded that they were arranged in an antiparallel fashion, more convincing evidence (19,20) appears to suggest that they are parallel to each.other, i.e. the two amino terminii are located at one end and the carboxy terminii at the other end.…”

Section: Dimer Formationmentioning

confidence: 99%

“…This prediction was corroborated by circular dichroic studies which showed that the c~-helical content of either L7 or L12 ranged between 45-60% (11, 2 !, 22), and that it could be increased to 75-80% in the presence of a helix promoting solvent (22). A higher value of about 76% has recently been suggested based on a more complete analysis of the circular dichroic spectrum (20). Gudkov et al (23) have determined the helicity of various fragments of L7.…”

Section: Physical Characteristicsmentioning

confidence: 99%

Chemistry and biology ofE. coli ribosomal protein L12

Brot

Weissbach

1981

Mol Cell Biochem

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E. coli ribosomal protein L12, because of its unique features, has been studied in more detail than perhaps any of the other ribosomal proteins. Unlike the other ribosomal proteins that are generally present in stoichiometric amounts, there are four copies of L12 per ribosome, some of which are acetylated on the N-terminal serine. The acetylated species, referred to as L7, has not been shown, as yet, to possess any different biological activity than L12. A specific enzyme that acetylates L12 to form L7, using acetyl-CoA as the acetyl donor, has been purified from E. coli extracts. L12 is also unique in that it does not contain cysteine, tryptophan, histidine, or tyrosine, is very acidic (pI: 4.85) and has a high content of ordered secondary structure (approximately 50%). The protein is normally found in solution as a dimer and also forms a tight complex with ribosomal protein L10. There are three methionine residues in L12, located in the N-terminal region of the protein, one or more of which are essential for biological activity. Oxidation of the methionines to methionine sulfoxide prevents dimer formation and inactivates the protein. The four copies of L12 are located in the crest region(s) of the 50S ribosomal subunit. There is good evidence that the soluble factors, such as IF-2, EF-Tu, EF-G and RF, interact with L12 on the ribosome during the process of protein synthesis. This interaction is essential for the proper functioning of each of the factors and for GTP hydrolysis associated with the individual partial reactions of protein synthesis. The L12 gene is located on an operon that contains the genes for L10 and beta beta' subunits of RNA polymerase at about 88 min on the bacterial chromosome. DNA-directed in vitro systems have been used to study the unique regulation of the expression of these genes. Autogenous regulation, translational control, and transcription attenuation are regulatory mechanisms that function to control the synthesis of these proteins.

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“…Figure 1 shows the structure and principle of the ICkit. The ICkit targets the ribosomal protein L7/L12 antigen, which is a component of the 50S ribosome and contains an amino acid sequence speci c to each bacterial species [20][21][22][23].…”

Section: Introductionmentioning

confidence: 99%

“…ICkit targets the ribosomal protein L7/L12 antigen, which is a component of the 50S ribosome and contains an amino acid sequence speci c to each bacterial species [20][21][22][23]. Sano et al and Ito et al reported the ICkit's utility in detecting the ribosomal protein L7/L12 in diagnosing Mycoplasma pneumonia and Legionella pneumonia [24,25].…”

Section: Introductionmentioning

confidence: 99%

Clinical Evaluation of a New Rapid Immunochromatographic Test for Detection of Bordetella Pertussis Antigen

Okada

Horikoshi

Nishimura

et al. 2021

Preprint

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A more rapid and less complicated test is required in clinical settings to diagnose pertussis. We need to detect Bordetella pertussis, which mainly causes pertussis, as early as possible, because pertussis is more likely to become severe in infants, and people around them can easily become the source of infection due to its strong infectivity. Nevertheless, methods that can detect B. pertussis rapidly and efficiently are lacking. Therefore, we developed a new immunochromatographic antigen kit (ICkit) for the early diagnosis of pertussis. The ICkit detects B. pertussis antigens in a nasopharyngeal swab without equipment and provides the result in about 15 min with a simple procedure. Additionally, a prospective study to evaluate the ICkit was conducted in 11 medical institutions, involving 195 cases with suspected pertussis. The sensitivity and specificity of the ICkit were 86.4% (19/22) and 97.1% (168/173), respectively, compared with the real-time polymerase chain reaction (rPCR). The ICkit detected the antigen in both children and adults. Furthermore, the ICkit detected the antigen until the 25th day from the onset of cough, when rPCR detected the antigen. Thus, the ICkit demonstrated a high correlation with rPCR and would help diagnose pertussis more rapidly and efficiently.

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Conformational stability of ribosomal protein L7/L12: effects of pH, temperature, and guanidinium chloride

Cited by 22 publications

References 48 publications

Protein Structures under Electrospray Conditions

Protein Structures under Electrospray Conditions

Chemistry and biology ofE. coli ribosomal protein L12

Clinical Evaluation of a New Rapid Immunochromatographic Test for Detection of Bordetella Pertussis Antigen

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