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Cloning and tissue distribution of human membrane-bound aminopeptidase P
R C Venema1, H Ju, R Zou
1Vascular Biology Center, Medical College of Georgia, Augusta 30912-2500, USA.
Abstract:
Complementary DNA clones encoding human membrane-bound aminopeptidase P (AmP) were isolated by reverse transcription-polymerase chain reaction (RT-PCR) of human kidney and lung poly (A)+ RNA. Comparison of the human AmP sequence to that of the pig shows significant evolutionary divergence with only 83% amino acid sequence identity between the two species. Northern hybridization analysis and RT-PCR suggests that the soluble and membrane-bound forms of human AmP are products of two distinct genes or, through alternative splicing, have different C-terminal sequences.
Insights
Researchers isolated complementary DNA clones for human membrane-bound aminopeptidase P (AmP). Human and pig AmP sequences show significant evolutionary divergence, suggesting distinct gene products or alternative splicing for soluble and membrane-bound forms.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Membrane-bound aminopeptidase P (AmP) plays crucial roles in biological processes.
- Understanding the genetic basis and evolutionary aspects of AmP is essential.
Purpose of the Study:
- To isolate and characterize complementary DNA (cDNA) clones encoding human membrane-bound aminopeptidase P (AmP).
- To investigate the evolutionary relationship between human and porcine AmP.
- To explore the genetic basis for soluble and membrane-bound forms of human AmP.
Main Methods:
- Complementary DNA (cDNA) cloning using reverse transcription-polymerase chain reaction (RT-PCR).
- Analysis of poly (A)+ RNA from human kidney and lung tissues.
- Sequence comparison between human and porcine AmP.
- Northern hybridization analysis.
Main Results:
- Successfully isolated cDNA clones for human membrane-bound aminopeptidase P (AmP).
- Human and porcine AmP sequences exhibit significant evolutionary divergence, with only 83% amino acid identity.
- Evidence suggests that soluble and membrane-bound human AmP forms arise from distinct genes or alternative splicing.
Conclusions:
- Human membrane-bound aminopeptidase P (AmP) has been characterized at the cDNA level.
- Significant evolutionary divergence exists between human and porcine AmP.
- The distinct forms of human AmP likely result from differential gene expression or alternative splicing mechanisms.