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J. Biol. Chem., Vol. 280, Issue 6, 4451-4461, February 11, 2005

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Peptidomics of Cpe^fat/fat Mouse Hypothalamus

EFFECT OF FOOD DEPRIVATION AND EXERCISE ON PEPTIDE LEVELS^*

Fa-Yun Che, Quan Yuan, Elena Kalinina, and Lloyd D. Fricker¶

From the Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461 and the Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China

Carboxypeptidase E is a major enzyme in the biosynthesis of numerous neuroendocrine peptides. Previously, we developed a technique for the isolation of neuropeptide-processing intermediates from mice that lack carboxypeptidase E activity (Cpe^fat/fat mice) due to a naturally occurring point mutation. In the present study, we used a differential labeling procedure with stable isotopic tags and mass spectrometry to quantitate the relative changes in a number of hypothalamic peptides in Cpe^fat/fat mice in two different paradigms that each cause an 10% decrease in body mass. One paradigm involved a 2-day fast under normal sedentary conditions (i.e. standard mouse cages); the other involved giving mice access to an exercise wheel for 4 weeks with free access to food. Approximately 50 peptides were detected in both studies, and over 80 peptides were detected in at least one of the two studies. Twenty-eight peptides were increased >50% by food deprivation, and some of these were increased by 2- to 3-fold. In contrast, only three peptides were increased >50% in the group with exercise wheels, and many peptides showed a slight 15–30% decrease upon exercise. Approximately one-half of the peptides detected in both studies were identified by tandem mass spectrometry. Peptides found to be elevated by food deprivation but not exercise included a number of fragments of proenkephalin, prothyrotropin-releasing hormone, secretogranin II, chromogranin B, and pro-SAAS. Taken together, the differential regulation of these peptides in the two paradigms suggests that the regulation is not due to the lower body weight but to the manner in which the paradigms achieved this lower body weight.

Received for publication, September 29, 2004 , and in revised form, November 24, 2004.

^* This work was supported primarily by National Institutes of Health Grant DK-67350 and by Grants DK-51271, DA-04494, and DA-17665 (to L. D. F.). The Laboratory for Macromolecular Analysis of the Albert Einstein College of Medicine is supported in part by Cancer Center Core Grant CA13330. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Dept. of Molecular Pharmacology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Tel.: 718-430-4225; Fax: 718-430-8954; E-mail: fricker{at}aecom.yu.edu.

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