CHARACTERIZATION OF HUMAN SURFACTANT APOPROTEINS

Project: Research project

Description

The purpose of this proposal is to characterize the apoprotein(s) of human
pulmonary surfactant using state-of-the-art electrophoretic techniques in
conjunction with immunochemical methods. This is the first step of a
detailed study of the intracellular and extracellular metabolism of the
apoproteins. Specifically, the various forms of surfactant apoprotein will
be identified in surfactant obtained from various human sources, including
the human lung cell line, A549, which synthesizes and secretes at least two
forms of surfactant apoprotein. Using these cells, the synthesis,
intracellular processing and extracellular processing will be examined by
electrophoresis, peptide mapping, isotopic labelling studies and
immunochemical methods in order to determine the relationship between these
proteins. Messenger RNA from these cells will then be translated in a
cell-free system in order to identify the primary translation product(s)
and determine whether the various forms of surfactant apoproteins are
derived from a single precursor molecule or are separate gene products.
From these studies, information will be gained about the properties of the
proteins that will be useful for their purification as well. This will
permit the testing of homogeneous preparations of various surfactant
apoprotein forms with natural and artificial surfactant preparations.
Well-characterized forms of surfactant apoproteins may provide more
specific endpoints for the study of hormonal regulation of fetal lung
muturation than phospholipid profile and morphology have offered to date.
Studies of the regulation of these proteins may then permit more effective
clinical detection and intervention in the prevention and treatment of
neonatal respiratory distress syndrome.
StatusFinished
Effective start/end date12/1/8411/30/86

Funding

  • National Institutes of Health
  • National Institutes of Health

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Apoproteins
Surface-Active Agents
Peptide Mapping
Phospholipids
Lung
Messenger RNA
Genes