Isolation and Characterization of Fatty Acid Binding Protein From Lactating Bovine Mammary Gland
Whetstone, Holly Diane
This item is only available for download by members of the University of Illinois community. Students, faculty, and staff at the U of I may log in with your NetID and password to view the item. If you are trying to access an Illinois-restricted dissertation or thesis, you can request a copy through your library's Inter-Library Loan office or purchase a copy directly from ProQuest.
Permalink
https://hdl.handle.net/2142/71727
Description
Title
Isolation and Characterization of Fatty Acid Binding Protein From Lactating Bovine Mammary Gland
Author(s)
Whetstone, Holly Diane
Issue Date
1985
Department of Study
Animal Science
Discipline
Dairy Science
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Agriculture, Animal Culture and Nutrition
Abstract
An intracellular protein which binds fatty acids had been found in a wide range of organisms and tissues. Its presence had not been documented in mammary tissue. The purpose of this study was to determine whether fatty acid binding protein (FABP) exists in bovine mammary tissue and if so, to isolate and characterize it. Mammary tissue was obtained at slaughter from five cows from the University dairy herd which were producing 5-25 kg milk/day. The tissue was homogenized and centrifuged to obtain the cytosolic and microsomal fractions. FABP was isolated from the cytosolic fraction by gel filtration on a Sephadex G-75 column followed by ion exchange chromatography on DEAE Sephadex A-25. FABP was eluted from the ion exchange column by .1 M NaCl. Molecular weight of FABP was estimated to be 12,000 by SDS polyacrylamide gel electrophoresis (PAGE). Two or three bands were seen on nondenaturing PAGE, two of which bound fatty acids. Activity of FABP was retained after incubation at 60 C for 15 min. FABP bound long chain fatty acids and their CoA derivatives but did not bind medium or short chain fatty acids. The affinity constant (Ka) of FABP for oleate was estimated to be 2 micromolar. The isolated FABP contained endogenous fatty acids, both covalently and noncovalently associated, which were primarily oleate, palmitate and stearate. The isoelectric point as determined by isoelectric focusing was about 5.6. Neither FABP nor albumin affected the activities of microsomal phosphatidic acid phosphatase, fatty acid:CoA ligase, or diacylglycerol acyltransferase. Fatty acid binding protein was not detected in skim milk or skim colostrum. This study demonstrated that mammary gland does possess a FABP which has properties similar to that found in other tissues.
Use this login method if you
don't
have an
@illinois.edu
email address.
(Oops, I do have one)
IDEALS migrated to a new platform on June 23, 2022. If you created
your account prior to this date, you will have to reset your password
using the forgot-password link below.
