Optimization of neuropeptide extraction from the mouse hypothalamus

Fa Yun Che; Xin Zhang; Iryna Berezniuk; Myrasol Callaway; Jihyeon Lim; Lloyd D. Fricker

doi:10.1021/pr060690r

Optimization of neuropeptide extraction from the mouse hypothalamus

Fa Yun Che, Xin Zhang, Iryna Berezniuk, Myrasol Callaway, Jihyeon Lim, Lloyd D. Fricker

Research output: Contribution to journal › Article › peer-review

58 Scopus citations

Abstract

Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70°C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides.

Original language	English (US)
Pages (from-to)	4667-4676
Number of pages	10
Journal	Journal of Proteome Research
Volume	6
Issue number	12
DOIs	https://doi.org/10.1021/pr060690r
State	Published - Dec 2007

Keywords

Carboxypeptidase D
Carboxypeptidase E
Peptide processing
Prohormone convertase

ASJC Scopus subject areas

General Chemistry
Biochemistry

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10.1021/pr060690r

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AU - Che, Fa Yun

AU - Zhang, Xin

AU - Berezniuk, Iryna

AU - Callaway, Myrasol

AU - Lim, Jihyeon

AU - Fricker, Lloyd D.

PY - 2007/12

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AB - Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70°C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides.

KW - Carboxypeptidase D

KW - Carboxypeptidase E

KW - Peptide processing

KW - Prohormone convertase

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Optimization of neuropeptide extraction from the mouse hypothalamus

Abstract

Keywords

ASJC Scopus subject areas

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