PMID- 1707846
OWN - NLM
STAT- MEDLINE
DCOM- 19910521
LR  - 20190707
IS  - 0378-1119 (Print)
IS  - 0378-1119 (Linking)
VI  - 98
IP  - 2
DP  - 1991 Feb 15
TI  - Synthesis and characterization of the Kunitz protease-inhibitor domain of the 
      beta-amyloid precursor protein.
PG  - 225-30
AB  - To understand the pathological process by which amyloid is deposited in 
      Alzheimer's disease, it is important to characterize the proteolytic processing 
      events of the beta-amyloid precursor protein (beta-APP) from which the 
      amyloid-forming fragment is excised. A potentially important component in 
      beta-APP processing is the 57-amino acid (aa) Kunitz serine protease inhibitor 
      (KPI) located within the extracellular domain of both the 751- and 770-aa 
      isoforms of beta-APP. We have synthesized DNA encoding the 57-aa KPI domain as a 
      necessary step in identifying the role of the protease inhibitor in beta-APP 
      processing and amyloid formation. A bacterial secretion system directed by the 
      alkaline phosphatase signal peptide of Escherichia coli linked to a synthetic 
      gene encoding KPI was used to produce soluble, extracellular recombinant KPI 
      (reKPI) protein. The reKPI protein was purified to homogeneity from bacterial 
      supernatants and was biochemically and biologically characterized. Complete aa 
      sequence analysis confirmed the fidelity of the reKPI, and fast-atom bombardment 
      mass-spectral analysis was used to document that reKPI was of the predicted Mr. 
      The reKPI is as active on a molar basis as the inhibitor-containing beta-APP when 
      assayed for inhibition of trypsin activity. Together these data suggest that 
      reKPI protein is properly folded and lacking in modified aa. Hence, this reKPI 
      will be an important reagent in gaining a better understanding of the role of the 
      KPI domain in beta-APP function and metabolism, as well as in the proteolytic 
      events involved in beta-amyloid formation.
FAU - Schilling, J
AU  - Schilling J
AD  - California Biotechnology Inc., Mountain View 94043.
FAU - Wang, Y
AU  - Wang Y
FAU - Lau, K
AU  - Lau K
FAU - Smith, L
AU  - Smith L
FAU - Cordell, B
AU  - Cordell B
LA  - eng
SI  - GENBANK/X06989
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (Amyloid beta-Peptides)
RN  - 0 (Amyloid beta-Protein Precursor)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (Protein Precursors)
RN  - 0 (Recombinant Proteins)
RN  - 9087-70-1 (Aprotinin)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - Alzheimer Disease/genetics
MH  - Amino Acid Sequence
MH  - Amyloid beta-Peptides/*genetics/pharmacology
MH  - Amyloid beta-Protein Precursor
MH  - Aprotinin/*genetics/isolation & purification/pharmacology
MH  - Base Sequence
MH  - Chromatography, High Pressure Liquid
MH  - Cloning, Molecular
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Molecular Sequence Data
MH  - Oligonucleotide Probes/chemical synthesis
MH  - Plasmids
MH  - Protein Precursors/*genetics/pharmacology
MH  - Recombinant Proteins/biosynthesis/isolation & purification/pharmacology
MH  - Restriction Mapping
MH  - Sequence Homology, Nucleic Acid
MH  - Trypsin/metabolism
EDAT- 1991/02/15 00:00
MHDA- 1991/02/15 00:01
CRDT- 1991/02/15 00:00
PHST- 1991/02/15 00:00 [pubmed]
PHST- 1991/02/15 00:01 [medline]
PHST- 1991/02/15 00:00 [entrez]
AID - 0378-1119(91)90177-D [pii]
AID - 10.1016/0378-1119(91)90177-d [doi]
PST - ppublish
SO  - Gene. 1991 Feb 15;98(2):225-30. doi: 10.1016/0378-1119(91)90177-d.