Reduction of Canine Plasminogen Leads to an Expanded Molecule which Precipitates

Kornblatt, Jack A.

doi:10.1371/journal.pone.0006196

Cited by 4 publications

(4 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There is one condition to date in which we have seen binding between native Str enolase, in solution, and dPgn. If dPgn is treated with reductant, such as dithiothreitol (DTT), the dPgn partially opens (as if it were a molten globule) and precipitates [49] . The precipitation can be monitored by light scattering at 280 nm.…”

Section: Resultsmentioning

confidence: 99%

“…“Pull down” reactions were of two types: (1) DTT induced precipitation and the course of the reaction followed at 280 nm have been described [49] and (2) adsorption of Str enolase and dPgn to phosphatidyl choline micelles. In the latter case, azolectin (2 mg/mL) was suspended in 10 mM potassium phosphate, 100 mM NaCl, pH 6.7 and sonicated until clear.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The Interaction of Canine Plasminogen with Streptococcus pyogenes Enolase: They Bind to One Another but What Is the Nature of the Structures Involved?

Kornblatt

Hancock

2011

PLoS ONE

Self Cite

View full text Add to dashboard Cite

For years it has been clear that plasminogen from different sources and enolase from different sources interact strongly. What is less clear is the nature of the structures required for them to interact. This work examines the interaction between canine plasminogen (dPgn) and Streptococcus pyogenes enolase (Str enolase) using analytical ultracentrifugation (AUC), surface plasmon resonance (SPR), fluorescence polarization, dynamic light scattering (DLS), isothermal titration calorimetry (ITC), and simple pull-down reactions. Overall, our data indicate that a non-native structure of the octameric Str enolase (monomers or multimers) is an important determinant of its surface-mediated interaction with host plasminogen. Interestingly, a non-native structure of plasminogen is capable of interacting with native enolase. As far as we can tell, the native structures resist forming stable mixed complexes.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Interaction of Canine Plasminogen with Streptococcus pyogenes Enolase: They Bind to One Another but What Is the Nature of the Structures Involved?

Kornblatt

Hancock

2011

PLoS ONE

Self Cite

View full text Add to dashboard Cite

show abstract

“…Because PrP Sc propagation was not further induced when plasminogen-supplemented PMCA included reducing agents, plasminogen does not appear to be the cofactor described by Supattapone's group. Furthermore, no concrete evidence indicates that it includes free sulfhydryl groups once secreted in its mature form (60), although plasminogen can acquire free sulfhydryl groups in vitro by interacting with cellular enzymes such as reductases (61). However, their observation that the reduced/oxidized state of the intramolecular disulfide bond of PrP is less critical in PrP conversion (31) coincides with our finding that PMCA conducted in the presence BME and DTT still allows basal amplification but abolishes the plasminogen-induced effect.…”

Section: Discussionmentioning

confidence: 99%

Plasminogen stimulates propagation of protease‐resistant prion protein in vitro

Mays

Ryou

2010

FASEB j.

View full text Add to dashboard Cite

To clarify the role of plasminogen as a cofactor for prion propagation, we conducted functional assays using a cell‐free prion protein (PrP) conversion assay termed protein misfolding cyclic amplification (PMCA) and prion‐infected cell lines. Here, we report that plasminogen stimulates propagation of the protease‐resistant scrapie PrP (PrPSc). Compared to control PMCA conducted without plasminogen, addition of plasminogen in PMCA using wild‐type brain material significantly increased PrP conversion, with an EC50 = ~56 nM. PrP conversion in PMCA was substantially less efficient with plasminogen‐deficient brain material than with wild‐type material. The activity stimulating PrP conversion was specific for plasminogen and conserved in its kringle domains. Such activity was abrogated by modification of plasminogen structure and interference of PrP‐plasminogen interaction. Kinetic analysis of PrPSc generation demonstrated that the presence of plasminogen in PMCA enhanced the PrPSc production rate to −0.97 U/µl/h and reduced turnover time to ~1 h compared to those (~0.4 U/µl/h and ~2.5 h) obtained without supplementation. Furthermore, as observed in PMCA, plasminogen and kringles promoted PrPSc propagation in ScN2a and Elk 21+ cells. Our results demonstrate that plasmino‐gen functions in stimulating conversion processes and represents the first cellular protein cofactor that enhances the hypothetical mechanism of prion propagation.—Mays, C. E., Ryou, C. Plasminogen stimulates propagation of protease‐resistant prion protein in vitro. FASEB J. 24, 5102–5112 (2010). http://www.fasebj.org

show abstract

“…Because the reducing agent is known to cause disturbances in the far UV region of the CD spectra, its concentration was kept below 1 mM as recommended in literature. [86] To prevent the reformation of the disulfide bond, all solvents were degased and flushed with a gas mixture of 5% hydrogen in nitrogen to apply anaerobic conditions. The CD spectra were recorded in the spectral range of 190 to 250 nm.…”

Section: Modification Of Zif268 With Artificial Dinuclear Amino Acidsmentioning

confidence: 99%

Development of Zinc-Finger-Based Artificial Restriction Endonucleases and Fluorescent Peptidyl Metal Sensors

Czerny¹

View full text Add to dashboard Cite

With more than thousand different members that have been discovered yet, zinc finger proteins are considered to be one of the most comprehensive classes of DNA-binding proteins found in nature. [9] This is due to their participation as DNA-binding domains in the modular structure of many eukaryotic and prokaryotic transcription factors. In this function, they are involved in the early steps of DNA transcription, an ubiquitous process that uses the genetic information by translating DNA sequences into corresponding messenger RNAs (mRNAs). Thereby, ZFPs recognize a specific promoter sequence, which is located upstream of the genetic code that should be transcribed by RNA-polymerases (Figure 1.1).Under retention of an uniform globular -structure, zinc fingers differ in their individual amino acid sequences to bind a large variety of promoter regions in a sequence-specific manner. This explains the high diversity of this protein family in the cellular environment and finally allows them to trigger transcriptional processes for a myriad of genes.

show abstract

Reduction of Canine Plasminogen Leads to an Expanded Molecule which Precipitates

Cited by 4 publications

References 46 publications

The Interaction of Canine Plasminogen with Streptococcus pyogenes Enolase: They Bind to One Another but What Is the Nature of the Structures Involved?

The Interaction of Canine Plasminogen with Streptococcus pyogenes Enolase: They Bind to One Another but What Is the Nature of the Structures Involved?

Plasminogen stimulates propagation of protease‐resistant prion protein in vitro

Development of Zinc-Finger-Based Artificial Restriction Endonucleases and Fluorescent Peptidyl Metal Sensors

Contact Info

Product

Resources

About