Proteolysis Assays With Conserved or Aminofluorescein‐Labeled Red Blood Cells.
| Title: | Proteolysis Assays With Conserved or Aminofluorescein‐Labeled Red Blood Cells. |
|---|---|
| Authors: | Al-Essa, Mohamed K.1 (AUTHOR) malessa@ju.edu.jo, Al-Qudah, Tamara1 (AUTHOR), Al Hadidi, Akram Kamal A.2 (AUTHOR), Alshubbak, Nida'a H.3 (AUTHOR), Siemianowicz, Krzysztof (AUTHOR) ksiem@mp.pl |
| Source: | BioMed Research International. 9/27/2024, Vol. 2024, p1-11. 11p. |
| Subject Terms: | *PEPTIDE analysis, *FLUORESCENT dyes, *PHENOMENOLOGICAL biology, *ERYTHROCYTES, *RESEARCH funding, *TRYPSIN, *CHEMICAL reagents, *BIOCHEMISTRY, *FLUORESCENT antibody technique, *CULTURE media (Biology), *CELL culture, *PROTEOLYTIC enzymes, *BIOLOGICAL assay, *RELIABILITY (Personality trait) |
| Abstract: | Backgrounds: Various physiological functions and reaction cascades, as well as disease progression in the living systems, are controlled by the activity of specific proteolytic enzymes. We conducted the study to evaluate protease activity by assessing peptide fragments from either conserved or labeled red blood cells (RBCs) with aminofluorescein (AF) in the reaction media. Methods: RBCs were incubated in media containing trypsin. Subsequently, the concentration of peptide fragments in the reaction media, resulted by the digestion with trypsin from conserved cells, was estimated by 3‐(4‐carboxybenzoyl)quinoline‐2‐carboxaldehyde (CBQCA) as an amine‐reactive fluorogenic reagent. In a second approach, we conjugated AF to the conserved RBCs and then exposed AF‐labeled RBCs to trypsin. This was followed by directly measuring the fluorescence intensity (FI) in the reaction media to estimate the concentration of AF‐labeled peptide fragments resulting from the enzyme's activity. Results: Show a concentration‐ and time‐dependent increase in FIs, reflecting the activity of trypsin as a proteolytic enzyme. The FIs increased significantly by 4 to 5 folds in samples treated with different enzyme concentrations, and by over 11 folds after 2 h incubation in media containing a 50 μL trypsin, as evidenced by CBQCA assays. Conclusion: These fast and affordable approaches could be applied with high reliability for the general estimation of protease activity in samples and customized for diagnostic purposes and prognostic evaluation in various diseases. [ABSTRACT FROM AUTHOR] |
| Copyright of BioMed Research International is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. 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: | Academic Search Complete |
| Full text is not displayed to guests. | Login for full access. |
| FullText | Links: – Type: pdflink Text: Availability: 1 CustomLinks: – Url: https://resolver.ebsco.com/c/xy5jbn/result?sid=EBSCO:a9h&genre=article&issn=23146133&ISBN=&volume=2024&issue=&date=20240927&spage=1&pages=1-11&title=BioMed Research International&atitle=Proteolysis%20Assays%20With%20Conserved%20or%20Aminofluorescein%E2%80%90Labeled%20Red%20Blood%20Cells.&aulast=Al-Essa%2C%20Mohamed%20K.&id=DOI:10.1155/2024/7919329 Name: Full Text Finder (for New FTF UI) (s8985755) Category: fullText Text: Find It @ SCU Libraries MouseOverText: Find It @ SCU Libraries |
|---|---|
| Header | DbId: a9h DbLabel: Academic Search Complete An: 179963150 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
| IllustrationInfo | |
| Items | – Name: Title Label: Title Group: Ti Data: Proteolysis Assays With Conserved or Aminofluorescein‐Labeled Red Blood Cells. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Al-Essa%2C+Mohamed+K%2E%22">Al-Essa, Mohamed K.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> malessa@ju.edu.jo</i><br /><searchLink fieldCode="AR" term="%22Al-Qudah%2C+Tamara%22">Al-Qudah, Tamara</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Al+Hadidi%2C+Akram+Kamal+A%2E%22">Al Hadidi, Akram Kamal A.</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Alshubbak%2C+Nida'a+H%2E%22">Alshubbak, Nida'a H.</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Siemianowicz%2C+Krzysztof%22">Siemianowicz, Krzysztof</searchLink> (AUTHOR)<i> ksiem@mp.pl</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22BioMed+Research+International%22">BioMed Research International</searchLink>. 9/27/2024, Vol. 2024, p1-11. 11p. – Name: Subject Label: Subject Terms Group: Su Data: *<searchLink fieldCode="DE" term="%22PEPTIDE+analysis%22">PEPTIDE analysis</searchLink><br />*<searchLink fieldCode="DE" term="%22FLUORESCENT+dyes%22">FLUORESCENT dyes</searchLink><br />*<searchLink fieldCode="DE" term="%22PHENOMENOLOGICAL+biology%22">PHENOMENOLOGICAL biology</searchLink><br />*<searchLink fieldCode="DE" term="%22ERYTHROCYTES%22">ERYTHROCYTES</searchLink><br />*<searchLink fieldCode="DE" term="%22RESEARCH+funding%22">RESEARCH funding</searchLink><br />*<searchLink fieldCode="DE" term="%22TRYPSIN%22">TRYPSIN</searchLink><br />*<searchLink fieldCode="DE" term="%22CHEMICAL+reagents%22">CHEMICAL reagents</searchLink><br />*<searchLink fieldCode="DE" term="%22BIOCHEMISTRY%22">BIOCHEMISTRY</searchLink><br />*<searchLink fieldCode="DE" term="%22FLUORESCENT+antibody+technique%22">FLUORESCENT antibody technique</searchLink><br />*<searchLink fieldCode="DE" term="%22CULTURE+media+%28Biology%29%22">CULTURE media (Biology)</searchLink><br />*<searchLink fieldCode="DE" term="%22CELL+culture%22">CELL culture</searchLink><br />*<searchLink fieldCode="DE" term="%22PROTEOLYTIC+enzymes%22">PROTEOLYTIC enzymes</searchLink><br />*<searchLink fieldCode="DE" term="%22BIOLOGICAL+assay%22">BIOLOGICAL assay</searchLink><br />*<searchLink fieldCode="DE" term="%22RELIABILITY+%28Personality+trait%29%22">RELIABILITY (Personality trait)</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Backgrounds: Various physiological functions and reaction cascades, as well as disease progression in the living systems, are controlled by the activity of specific proteolytic enzymes. We conducted the study to evaluate protease activity by assessing peptide fragments from either conserved or labeled red blood cells (RBCs) with aminofluorescein (AF) in the reaction media. Methods: RBCs were incubated in media containing trypsin. Subsequently, the concentration of peptide fragments in the reaction media, resulted by the digestion with trypsin from conserved cells, was estimated by 3‐(4‐carboxybenzoyl)quinoline‐2‐carboxaldehyde (CBQCA) as an amine‐reactive fluorogenic reagent. In a second approach, we conjugated AF to the conserved RBCs and then exposed AF‐labeled RBCs to trypsin. This was followed by directly measuring the fluorescence intensity (FI) in the reaction media to estimate the concentration of AF‐labeled peptide fragments resulting from the enzyme's activity. Results: Show a concentration‐ and time‐dependent increase in FIs, reflecting the activity of trypsin as a proteolytic enzyme. The FIs increased significantly by 4 to 5 folds in samples treated with different enzyme concentrations, and by over 11 folds after 2 h incubation in media containing a 50 μL trypsin, as evidenced by CBQCA assays. Conclusion: These fast and affordable approaches could be applied with high reliability for the general estimation of protease activity in samples and customized for diagnostic purposes and prognostic evaluation in various diseases. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of BioMed Research International is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. 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.) |
| PLink | https://login.libproxy.scu.edu/login?url=https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&scope=site&db=a9h&AN=179963150 |
| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1155/2024/7919329 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 11 StartPage: 1 Subjects: – SubjectFull: PEPTIDE analysis Type: general – SubjectFull: FLUORESCENT dyes Type: general – SubjectFull: PHENOMENOLOGICAL biology Type: general – SubjectFull: ERYTHROCYTES Type: general – SubjectFull: RESEARCH funding Type: general – SubjectFull: TRYPSIN Type: general – SubjectFull: CHEMICAL reagents Type: general – SubjectFull: BIOCHEMISTRY Type: general – SubjectFull: FLUORESCENT antibody technique Type: general – SubjectFull: CULTURE media (Biology) Type: general – SubjectFull: CELL culture Type: general – SubjectFull: PROTEOLYTIC enzymes Type: general – SubjectFull: BIOLOGICAL assay Type: general – SubjectFull: RELIABILITY (Personality trait) Type: general Titles: – TitleFull: Proteolysis Assays With Conserved or Aminofluorescein‐Labeled Red Blood Cells. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Al-Essa, Mohamed K. – PersonEntity: Name: NameFull: Al-Qudah, Tamara – PersonEntity: Name: NameFull: Al Hadidi, Akram Kamal A. – PersonEntity: Name: NameFull: Alshubbak, Nida'a H. – PersonEntity: Name: NameFull: Siemianowicz, Krzysztof IsPartOfRelationships: – BibEntity: Dates: – D: 27 M: 09 Text: 9/27/2024 Type: published Y: 2024 Identifiers: – Type: issn-print Value: 23146133 Numbering: – Type: volume Value: 2024 Titles: – TitleFull: BioMed Research International Type: main |
| ResultId | 1 |