Polymeric fluorescent heparin as one-step FRET substrate of human heparanase.
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| Title: | Polymeric fluorescent heparin as one-step FRET substrate of human heparanase. |
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| Authors: | Sistla, Jyothi C.1,2, Morla, Shravan1,2, Alabbas, Al-Humaidi B.1,2, Kalathur, Ravi C.3, Sharon, Chetna4, Patel, Bhaumik B.4,5, Desai, Umesh R.1,2 urdesai@vcu.edu |
| Source: | Carbohydrate Polymers. Feb2019, Vol. 205, p385-391. 7p. |
| Subjects: | Fluorescence resonance energy transfer, Polymeric composites, Heparin, Heparanase, Cell lines |
| Abstract: | Highlights • Development of polymeric heparin as a FRET substrate of heparanase. • The FRET substrate enables rapid, one-step screening of heparanase inhibitors. • The FRET substrate enables monitoring of heparanase activity in biological media. Abstract Heparanase, an endo-β- D -glucuronidase, cleaves cell surface and extracellular matrix heparan sulfate (HS) chains and plays important roles in cellular growth and metastasis. Heparanase assays reported to-date are labor intensive, complex and/or expensive. A simpler assay is critically needed to understand the myriad roles of heparanase. We reasoned that fluorescent heparin could serve as an effective probe of heparanase levels. Following synthesis and screening, a heparin preparation labeled with DABCYL and EDANS was identified, which exhibited a characteristic increase in signal following cleavage by human heparanase. This work describes the synthesis of this heparin substrate, its kinetic and spectrofluorometric properties, optimization of the heparanase assay, use of the assay in inhibitor screening, and elucidation of the state of heparanase in different cell lines. Our FRET-based assay is much simpler and more robust than all assays reported in the literature as well as a commercially available kit. [ABSTRACT FROM AUTHOR] |
| Copyright of Carbohydrate Polymers is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 132992575 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Polymeric fluorescent heparin as one-step FRET substrate of human heparanase. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Sistla%2C+Jyothi+C%2E%22">Sistla, Jyothi C.</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Morla%2C+Shravan%22">Morla, Shravan</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Alabbas%2C+Al-Humaidi+B%2E%22">Alabbas, Al-Humaidi B.</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Kalathur%2C+Ravi+C%2E%22">Kalathur, Ravi C.</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Sharon%2C+Chetna%22">Sharon, Chetna</searchLink><relatesTo>4</relatesTo><br /><searchLink fieldCode="AR" term="%22Patel%2C+Bhaumik+B%2E%22">Patel, Bhaumik B.</searchLink><relatesTo>4,5</relatesTo><br /><searchLink fieldCode="AR" term="%22Desai%2C+Umesh+R%2E%22">Desai, Umesh R.</searchLink><relatesTo>1,2</relatesTo><i> urdesai@vcu.edu</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Carbohydrate+Polymers%22">Carbohydrate Polymers</searchLink>. Feb2019, Vol. 205, p385-391. 7p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Fluorescence+resonance+energy+transfer%22">Fluorescence resonance energy transfer</searchLink><br /><searchLink fieldCode="DE" term="%22Polymeric+composites%22">Polymeric composites</searchLink><br /><searchLink fieldCode="DE" term="%22Heparin%22">Heparin</searchLink><br /><searchLink fieldCode="DE" term="%22Heparanase%22">Heparanase</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+lines%22">Cell lines</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Highlights • Development of polymeric heparin as a FRET substrate of heparanase. • The FRET substrate enables rapid, one-step screening of heparanase inhibitors. • The FRET substrate enables monitoring of heparanase activity in biological media. Abstract Heparanase, an endo-β- D -glucuronidase, cleaves cell surface and extracellular matrix heparan sulfate (HS) chains and plays important roles in cellular growth and metastasis. Heparanase assays reported to-date are labor intensive, complex and/or expensive. A simpler assay is critically needed to understand the myriad roles of heparanase. We reasoned that fluorescent heparin could serve as an effective probe of heparanase levels. Following synthesis and screening, a heparin preparation labeled with DABCYL and EDANS was identified, which exhibited a characteristic increase in signal following cleavage by human heparanase. This work describes the synthesis of this heparin substrate, its kinetic and spectrofluorometric properties, optimization of the heparanase assay, use of the assay in inhibitor screening, and elucidation of the state of heparanase in different cell lines. Our FRET-based assay is much simpler and more robust than all assays reported in the literature as well as a commercially available kit. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Carbohydrate Polymers is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.carbpol.2018.10.071 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 7 StartPage: 385 Subjects: – SubjectFull: Fluorescence resonance energy transfer Type: general – SubjectFull: Polymeric composites Type: general – SubjectFull: Heparin Type: general – SubjectFull: Heparanase Type: general – SubjectFull: Cell lines Type: general Titles: – TitleFull: Polymeric fluorescent heparin as one-step FRET substrate of human heparanase. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Sistla, Jyothi C. – PersonEntity: Name: NameFull: Morla, Shravan – PersonEntity: Name: NameFull: Alabbas, Al-Humaidi B. – PersonEntity: Name: NameFull: Kalathur, Ravi C. – PersonEntity: Name: NameFull: Sharon, Chetna – PersonEntity: Name: NameFull: Patel, Bhaumik B. – PersonEntity: Name: NameFull: Desai, Umesh R. IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 02 Text: Feb2019 Type: published Y: 2019 Identifiers: – Type: issn-print Value: 01448617 Numbering: – Type: volume Value: 205 Titles: – TitleFull: Carbohydrate Polymers Type: main |
| ResultId | 1 |
