Human cystatin D: cDNA cloning, characterization of the E. coli expressed inhibitor, and identification of the native protein in saliva
Freije, José P ; Balbin, Milagros ; Abrahamson, Magnus LU ; Velasco, Gloria ; Dalboge, Henrik ; Grubb, Anders LU
and Lopez-Otin, Carlos
(1993)
In Journal of Biological Chemistry
268(21).
p.15737-15744
- Abstract
- A cDNA coding for cystatin D, a human member of the cystatin protein family, has been cloned after specific amplification of reverse- transcribed parotid gland RNA. After replacing the segment encoding the putative 20-residue signal peptide with one encoding the Escherichia coli OmpA leader sequence, the cDNA was expressed in E. coli. The isolated recombinant protein exhibited Ki values of 1.2 nM and > 1 microM for papain and cathepsin B, respectively. An antiserum raised against recombinant cystatin D recognized a protein in human saliva with electrophoretical mobility identical to that of the recombinant protein. Immunoenzymatic analysis revealed that this cysteine proteinase inhibitor is present in human saliva and tears at... (More)
- A cDNA coding for cystatin D, a human member of the cystatin protein family, has been cloned after specific amplification of reverse- transcribed parotid gland RNA. After replacing the segment encoding the putative 20-residue signal peptide with one encoding the Escherichia coli OmpA leader sequence, the cDNA was expressed in E. coli. The isolated recombinant protein exhibited Ki values of 1.2 nM and > 1 microM for papain and cathepsin B, respectively. An antiserum raised against recombinant cystatin D recognized a protein in human saliva with electrophoretical mobility identical to that of the recombinant protein. Immunoenzymatic analysis revealed that this cysteine proteinase inhibitor is present in human saliva and tears at concentrations of 3.8 and 0.5 mg/liter, respectively, while it was not detected in seminal plasma, blood plasma, milk, or cerebrospinal fluid. Cystatin D purified from human saliva by immunosorption displayed a heterogeneous N-terminal end, with sequences starting at residues 5, 7, 9, and 11 of the predicted N-terminal portion of the mature protein. On the basis of structural and functional properties, cystatin D represents a novel cysteine proteinase inhibitor possibly playing a protective role against proteinases present in the oral cavity. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1107645
- author
- Freije, José P
; Balbin, Milagros
; Abrahamson, Magnus
LU
; Velasco, Gloria
; Dalboge, Henrik
; Grubb, Anders
LU
and Lopez-Otin, Carlos
- organization
- publishing date
- 1993
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 268
- issue
- 21
- pages
- 15737 - 15744
- publisher
- American Society for Biochemistry and Molecular Biology
- ISSN
- 1083-351X
- language
- English
- LU publication?
- yes
- id
- 23272562-6fe1-4bf2-9eeb-f97b7213ac33 (old id 1107645)
- alternative location
- http://www.jbc.org/cgi/reprint/268/21/15737
- date added to LUP
- 2016-04-01 11:43:36
- date last changed
- 2018-11-21 19:59:34
@article{23272562-6fe1-4bf2-9eeb-f97b7213ac33,
abstract = {{A cDNA coding for cystatin D, a human member of the cystatin protein family, has been cloned after specific amplification of reverse- transcribed parotid gland RNA. After replacing the segment encoding the putative 20-residue signal peptide with one encoding the Escherichia coli OmpA leader sequence, the cDNA was expressed in E. coli. The isolated recombinant protein exhibited Ki values of 1.2 nM and > 1 microM for papain and cathepsin B, respectively. An antiserum raised against recombinant cystatin D recognized a protein in human saliva with electrophoretical mobility identical to that of the recombinant protein. Immunoenzymatic analysis revealed that this cysteine proteinase inhibitor is present in human saliva and tears at concentrations of 3.8 and 0.5 mg/liter, respectively, while it was not detected in seminal plasma, blood plasma, milk, or cerebrospinal fluid. Cystatin D purified from human saliva by immunosorption displayed a heterogeneous N-terminal end, with sequences starting at residues 5, 7, 9, and 11 of the predicted N-terminal portion of the mature protein. On the basis of structural and functional properties, cystatin D represents a novel cysteine proteinase inhibitor possibly playing a protective role against proteinases present in the oral cavity.}},
author = {{Freije, José P and Balbin, Milagros and Abrahamson, Magnus and Velasco, Gloria and Dalboge, Henrik and Grubb, Anders and Lopez-Otin, Carlos}},
issn = {{1083-351X}},
language = {{eng}},
number = {{21}},
pages = {{15737--15744}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Human cystatin D: cDNA cloning, characterization of the E. coli expressed inhibitor, and identification of the native protein in saliva}},
url = {{http://www.jbc.org/cgi/reprint/268/21/15737}},
volume = {{268}},
year = {{1993}},
}