Recombinant Cyclophilins Lack Nuclease Activity

Manteca, Ángel; Sánchez, Jesús

doi:10.1128/jb.186.18.6325-6326.2004

Cited by 9 publications

(8 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The S. antibioticus cytosolic cyclophilin characterized in this work was proposed in a former report to be also a nuclease [33], as analogously reported for NUC-18, a thymocyte nuclease which was found to share amino terminal end sequence homology with proteins belonging to the cyclophilin family and which was proposed to play the key role in glucocorticoidstimulated apoptosis [34]. However, an analysis of the San-Cyp18 recombinant protein has shown the absence of such cyclophilin-associated nuclease activity [53].…”

Section: Discussionsupporting

confidence: 76%

Cloning and characterization of a Streptomyces antibioticus ATCC11891 cyclophilin related to Gram negative bacteria cyclophilins

et al. 2004

Self Cite

View full text Add to dashboard Cite

Cyclophilins are folding helper enzymes and represent a family of the enzyme class of peptidyl-prolyl cis-trans isomerases. Here, we report the molecular cloning and biochemical characterization of SanCyp18, an 18-kDa cyclophilin from Streptomyces antibioticus ATCC11891 located in the cytoplasm and constitutively expressed during development. Amino acid sequence analysis revealed a much higher homology to cyclophilins from Gram negative bacteria than to known cyclophilins from Streptomyces or other Gram positive bacteria. SanCyp18 is inhibited weakly by CsA, with a K i value of 21 lM, similar to cyclophilins from Gram negative bacteria. However, this value is more than 20-fold higher than the K i values reported for cyclophilins from other Gram positive bacteria, which makes SanCyp18 unique within this group. The presence of SanCyp18 in Streptomyces is likely due to horizontal gene transmission from Gram-negative bacteria to Streptomyces.

show abstract

Section: Discussionsupporting

confidence: 76%

Cloning and characterization of a Streptomyces antibioticus ATCC11891 cyclophilin related to Gram negative bacteria cyclophilins

et al. 2004

Self Cite

View full text Add to dashboard Cite

show abstract

“…13A) and one of the helices (aa. [136][137][138][139][140][141][142][143][144][145][146] and part of the b-barrel of CypA (29). Binding kinetics between recombinant AIF (unspecified construct) and human CypA was investigated by surface plasmon resonance in the presence of NADPH (253).…”

Section: Cyclophilin a Cyclophilins Possess A Peptidyl Prolyl Isomentioning

confidence: 99%

“…Cyclophilin A. Presently, it is unclear whether cyclophilins possess or lack nucleolytic activities (145,158). Nonetheless, recombinant CypA was demonstrated to cooperate with AIF in vitro in chromatinolysis and plasmid DNA degradation (29).…”

Section: Sevrioukovamentioning

confidence: 99%

Apoptosis-Inducing Factor: Structure, Function, and Redox Regulation

Sevrioukova

2011

Antioxidants & Redox Signaling

291

276

View full text Add to dashboard Cite

Apoptosis-inducing factor (AIF) is a flavin adenine dinucleotide-containing, NADH-dependent oxidoreductase residing in the mitochondrial intermembrane space whose specific enzymatic activity remains unknown. Upon an apoptotic insult, AIF undergoes proteolysis and translocates to the nucleus, where it triggers chromatin condensation and large-scale DNA degradation in a caspase-independent manner. Besides playing a key role in execution of caspase-independent cell death, AIF has emerged as a protein critical for cell survival. Analysis of in vivo phenotypes associated with AIF deficiency and defects, and identification of its mitochondrial, cytoplasmic, and nuclear partners revealed the complexity and multilevel regulation of AIF-mediated signal transduction and suggested an important role of AIF in the maintenance of mitochondrial morphology and energy metabolism. The redox activity of AIF is essential for optimal oxidative phosphorylation. Additionally, the protein is proposed to regulate the respiratory chain indirectly, through assembly and/or stabilization of complexes I and III. This review discusses accumulated data with respect to the AIF structure and outlines evidence that supports the prevalent mechanistic view on the apoptogenic actions of the flavoprotein, as well as the emerging concept of AIF as a redox sensor capable of linking NAD(H)-dependent metabolic pathways to apoptosis.

show abstract

“…Our results unequivocally indicate that CypB acts as a nuclease at supra physiological concentrations (>50 nM). Previous groups have debated CypB's nuclease activity beginning in 1997 with Montague and colleagues [14] and continuing through the early 2000s with Manteca and Sanchez [16] . Through our two‐pronged approach encompassing highly sensitive nucleic acid‐based electrochemical assays and mass spectrometry, here we demonstrate that ≥98 % pure recombinant human CypB has nuclease activity against single stranded and double stranded DNA, as well as against F‐RNA.…”

Section: Discussionmentioning

confidence: 75%

Human Cyclophilin B Nuclease Activity Revealed via Nucleic Acid‐Based Electrochemical Sensors

et al. 2022

View full text Add to dashboard Cite

Human cyclophilin B (CypB) is oversecreted by pancreatic cancer cells, making it a potential biomarker for early-stage disease diagnosis. Our group is motivated to develop aptamer-based assays to measure CypB levels in biofluids. However, human cyclophilins have been postulated to have collateral nuclease activity, which could impede the use of aptamers for CypB detection. To establish if CypB can hydrolyze electrode-bound nucleic acids, we used ultrasensitive electrochemical sensors to measure CypB's hydrolytic activity. Our sensors use ssDNA and dsDNA in the biologically predominant D-DNA form, and in the nuclease resistant L-DNA form. Challenging such sensors with CypB and control proteins, we unequivocally demonstrate that CypB can cleave nucleic acids. To our knowledge, this is the first study to use electrochemical biosensors to reveal the hydrolytic activity of a protein that is not known to be a nuclease. Future development of CypB bioassays will require the use of nuclease-resistant aptamer sequences.

show abstract

Recombinant Cyclophilins Lack Nuclease Activity

Cited by 9 publications

References 13 publications

Cloning and characterization of a Streptomyces antibioticus ATCC11891 cyclophilin related to Gram negative bacteria cyclophilins

Cloning and characterization of a Streptomyces antibioticus ATCC11891 cyclophilin related to Gram negative bacteria cyclophilins

Apoptosis-Inducing Factor: Structure, Function, and Redox Regulation

Human Cyclophilin B Nuclease Activity Revealed via Nucleic Acid‐Based Electrochemical Sensors

Contact Info

Product

Resources

About