Identification of enzyme-sensitive cleavage sites of beta endorphin (beta- END) using serum and inflamed tissue

Herath, Dilanthi, Hewavitharana, Amitha K., Cabot, Peter J. and Shaw, P. Nicholas (2011). Identification of enzyme-sensitive cleavage sites of beta endorphin (beta- END) using serum and inflamed tissue. In: 22nd American Peptide Society Symposium. American Peptide Symposium (22nd, 2011), San Diego, CA, U.S.A., (509-509). 25-30 June 2011. doi:10.1002/bip.21670

Author Herath, Dilanthi
Hewavitharana, Amitha K.
Cabot, Peter J.
Shaw, P. Nicholas
Title of paper Identification of enzyme-sensitive cleavage sites of beta endorphin (beta- END) using serum and inflamed tissue
Formatted title
Identification of enzyme-sensitive cleavage sites of beta-endorphin (β -END) using serum and inflamed tissue
Conference name American Peptide Symposium (22nd, 2011)
Conference location San Diego, CA, U.S.A.
Conference dates 25-30 June 2011
Proceedings title 22nd American Peptide Society Symposium   Check publisher's open access policy
Journal name Peptide Science   Check publisher's open access policy
Place of Publication Hoboken, NJ, U.S.A.
Publisher John Wiley & Sons
Publication Year 2011
Sub-type Published abstract
DOI 10.1002/bip.21670
ISSN 0006-3525
Volume 96
Issue 4
Start page 509
End page 509
Total pages 1
Language eng
Formatted Abstract/Summary
A study was undertaken to investigate the enzyme-sensitive cleavage sites of β-END (1-31) in serum and in peripheral inflamed tissue. Inflammatory rat paw tissue was homogenized either with PBS (pH 7 .4) or MES (pH 5.5) buffer (1 mg tissue: 10ml buffer) and centrifuged. Rat β-END ( 1-31) was added to the resultant supenatants of the inflamed tissue extracts (to a final concentration of 0.1 mM) and to rat serum (to a final concentration of 74 μM). All samples were incubated at 37°C. Following incubation periods ranging from 0 min to 120 min, a sample (100 μL) was withdrawn and the protein precipitated by the addition of trichloroacetic acid. Each sample was centrifuged and resultant supernatant was analyzed with LC-MS (total ion current spectra) and LC-MS- MS (fragmented ion spectra).

The primary enzymatic cleavage of β-END in rat serum occurs mainly at 16- 17-18-19-20 region that represents the amino acids Thr-Leu-Phe-Lys-Asn. For all primary fragments, the cleavage sites observed at pH 7.4 with inflamed tissue (except Lys-Ser at 9-10 position) were similar to those observed with rat serum. At pH 5.5, in addition to the cleavage sites detected in serum and in inflamed tissue at pH 7 .4, other sites were detected. These fragments were produced as a result of hydrolysis of β-END ( 1-31) by a variety of enzymes in the serum and inflammatory tissue. It is possible that the additional cleavage sites detected at pH 5.5 may be due to the enhanced enzymatic activity generated by the acidic environment of the inflammatory matrix.
Q-Index Code EX
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published under Poster Abstracts as Abstract "YI -P 350".

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