Document Detail


Comparison of brain and pituitary immunoreactive prolactin by peptide mapping and lectin affinity chromatography.
MedLine Citation:
PMID:  2791995     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Immunoreactive PRL (IR-PRL) has been identified in many areas of the rat brain. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses we have recently shown that the primary IR-PRL protein in the rat brain has an apparent mol wt (Mr) of 24,000, which was identical to that of pituitary PRL. In these studies, brain and pituitary 24,000 Mr PRL were compared by peptide mapping and lectin chromatography. PRL-enriched fractions were prepared from the pituitary, hypothalamus, hippocampus, and pons-medulla and labeled with 125I. This material was further purified by immunoprecipitation, and immunopurified 24,000 Mr PRL was isolated by sodium dodecyl sulfate-gel electrophoresis. Cleavage of 125I-labeled 24,000 Mr IR-PRL prepared from the pituitary and the three brain regions with chymotrypsin resulted in identical peptide maps with two primary labeled peptide fragments (5,500 and 6,000 Mr), and approximately five less intense fragments. Similarly, trypsin cleavage of brain and pituitary 24,000 Mr IR-PRL resulted in the production of two major fragments (6,200, and 5,200 Mr), and three less intense fragments. Cleavage of the 24,000 Mr IR-PRL with Staphylococcus V8 protease resulted in identical fragment patterns with two primary peptide fragments (14,000 and 6,200 Mr). When the pituitary and brain 24,000 Mr PRL were applied to Concanavalin-A-Sepharose columns no 24,000 Mr PRL was absorbed. The similarity of the peptide fragments obtained from the cleavage of the 24,000 Mr IR-PRL from the brain and pituitary clearly indicate that the IR-PRL found in the brain has an amino acid sequence that shares a high degree of structural homology with pituitary PRL.
Authors:
W J DeVito
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Endocrinology     Volume:  125     ISSN:  0013-7227     ISO Abbreviation:  Endocrinology     Publication Date:  1989 Nov 
Date Detail:
Created Date:  1989-11-22     Completed Date:  1989-11-22     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0375040     Medline TA:  Endocrinology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2439-44     Citation Subset:  AIM; IM    
Affiliation:
Division of Endocrinology, University of Massachusetts Medical School, Worcester 01655.
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MeSH Terms
Descriptor/Qualifier:
Animals
Brain Chemistry*
Chromatography, Affinity / methods
Concanavalin A
Electrophoresis, Polyacrylamide Gel
Hypophysectomy
Male
Molecular Weight
Organ Specificity
Peptide Mapping
Pituitary Gland / analysis*
Prolactin / analysis*,  isolation & purification
Rats
Grant Support
ID/Acronym/Agency:
DK-32520/DK/NIDDK NIH HHS; NS-26343/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
11028-71-0/Concanavalin A; 9002-62-4/Prolactin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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