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Processing of the native nerve growth factor precursor to form biologically active nerve growth factor
R H Edwards1, M J Selby, P D Garcia
1Departments of Biochemistry and Biophysics, University of California, San Francisco 94143.
The Journal of Biological Chemistry
|May 15, 1988
Summary
Mouse submaxillary gland studies reveal that the gamma-subunit of nerve growth factor (NGF) cleaves the beta-NGF precursor into biologically active beta-NGF. This enzymatic processing, along with trypsin, is crucial for NGF maturation and function.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Beta-nerve growth factor (beta-NGF) in mouse submaxillary glands forms complexes with kallikrein family serine proteases (alpha- and gamma-subunits).
- Understanding the processing of beta-NGF is essential for its biological activity.
Purpose of the Study:
- To investigate the role of the gamma-subunit in cleaving the beta-NGF precursor.
- To determine the conditions required for producing biologically active beta-NGF.
Main Methods:
- Recombinant vaccinia virus was used to produce the beta-NGF precursor in mammalian cells.
- Stoichiometric quantities of the gamma-subunit and catalytic quantities of trypsin were used to cleave the precursor.
Main Results:
- The gamma-subunit effectively cleaved the beta-NGF precursor to yield active beta-NGF.
- Trypsin also produced native beta-NGF, indicating alternative processing pathways.
- Precursor conformation was critical for proper cleavage.
- The resulting beta-NGF exhibited at least 10-fold greater biological activity compared to its precursor.
Conclusions:
- The gamma-subunit of NGF plays a significant role in the maturation of beta-NGF.
- Enzymatic cleavage by the gamma-subunit or trypsin is essential for generating biologically active beta-NGF.
- The conformational state of the precursor directly impacts the efficiency of its processing and subsequent biological activity.