Optimization of neuropeptide extraction from the mouse hypothalamus

被引：47

作者：

Che, Fa-Yun ^{[1
]}

Zhang, Xin ^{[1
]}

Berezniuk, Iryna ^{[1
]}

Callaway, Myrasol ^{[1
]}

Lim, Jihyeon ^{[1
]}

Fricker, Lloyd D. ^{[1
]}

机构：

[1] Albert Einstein Coll Med, Dept Mol Pharmacol, Bronx, NY 10461 USA

来源：

JOURNAL OF PROTEOME RESEARCH | 2007年 / 6卷 / 12期

关键词：

peptide processing; carboxypeptidase E; carboxypeptidase D; prohormone convertase;

D O I：

10.1021/pr060690r

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 degrees 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.

引用

页码：4667 / 4676

页数：10

共 47 条

[1]

ALLEN G, 1989, SEQUENCING PROTEINS, P73

[2] Peptidomics [J].

Baggerman, G ;

Verleyen, P ;

Clynen, E ;

Huybrechts, J ;

De Loof, A ;

Schoofs, L .

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2004, 803 (01) :3-16

[3]

Bennett HPJ, 1991, PEPTIDE BIOSYNTHESIS, P111

[4] Defective prodynorphin processing in mice lacking prohormone convertase PC2 [J].

Berman, Y ;

Mzhavia, N ;

Polonskaia, A ;

Furuta, M ;

Steiner, DF ;

Pintar, JE ;

Devi, LA .

JOURNAL OF NEUROCHEMISTRY, 2000, 75 (04) :1763-1770

[5]

CHARD T, 1987, INTRO RADIOIMMUNOASS, P1

[6] Quantitative peptidomics in mice - Effect of cocaine treatment [J].

Che, FY ;

Vathy, I ;

Fricker, LD .

JOURNAL OF MOLECULAR NEUROSCIENCE, 2006, 28 (03) :265-275

[7] Identification of peptides from brain and pituitary of Cpefat/Cpefat mice [J].

Che, FY ;

Yan, L ;

Li, H ;

Mzhavia, N ;

Devi, LA ;

Fricker, LD .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (17) :9971-9976

[8] Quantitative neuropeptidomics of microwave-irradiated mouse brain and pituitary [J].

Che, FY ;

Lim, J ;

Pan, H ;

Biswas, R ;

Fricker, LD .

MOLECULAR & CELLULAR PROTEOMICS, 2005, 4 (09) :1391-1405

[9] Relative quantitation of peptides in wild-type and Cpefat/fat mouse pituitary using stable isotopic tags and mass spectrometry [J].

Che, FY ;

Biswas, R ;

Fricker, LD .

JOURNAL OF MASS SPECTROMETRY, 2005, 40 (02) :227-237

[10] Quantitative peptidomics of mouse pituitary: comparison of different stable isotopic tags [J].

Che, FY ;

Fricker, LD .

JOURNAL OF MASS SPECTROMETRY, 2005, 40 (02) :238-249

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