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Molecular cloning and sequence analysis of cDNA encoding human prostatic acid phosphatase
P Vihko1, P Virkkunen, P Henttu
1Biocenter, University of Oulu, Finland.
FEBS Letters
|August 29, 1988
Summary
Researchers isolated human prostatic acid phosphatase (PAP) cDNA, revealing a 354-amino acid protein. Further analysis is ongoing to determine if PAP belongs to a gene family.
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- Human prostatic acid phosphatase (PAP) is an enzyme involved in prostate function.
- Understanding the genetic basis of PAP is crucial for research into prostate health and disease.
Purpose of the Study:
- To isolate and characterize the complementary DNA (cDNA) encoding human prostatic acid phosphatase (PAP).
- To determine the full coding sequence and protein structure of PAP.
- To investigate potential relationships between PAP and other phosphatases.
Main Methods:
- Isolation of lambda gt11 clones from human prostatic cDNA libraries using immunoscreening with polyclonal antisera.
- DNA sequencing of overlapping cDNA clones to determine the composite sequence.
- Direct protein sequencing of the mature PAP protein and its fragments.
- RNA blot analysis to determine mRNA size.
Main Results:
- Successfully isolated and sequenced cDNA clones encoding human PAP, a 354-residue protein (41,126 Da) with a 32-amino acid signal peptide.
- Confirmed the predicted protein sequence through direct amino-terminal and fragment sequencing.
- Identified an alu-type repetitive sequence in the 3'-untranslated region of PAP cDNA, similar to that found in human placental alkaline phosphatase.
- Observed variations at the 5'-ends of some cDNA clones and confirmed a 3.3 kb mRNA size.
Conclusions:
- The complete coding region for human PAP has been identified, providing a foundation for further functional studies.
- PAP shows no significant sequence homology to other known proteins, suggesting unique evolutionary origins.
- The presence of repetitive sequences and variations in cDNA clones warrants further investigation into the possibility of PAP belonging to a gene family, analogous to alkaline phosphatases.