Cystatin F is a glycosylated human low molecular weight cysteine proteinase inhibitor
Ni, J ; Alvarez Fernandez, Marcia LU ; Danielsson, Lena ; Chillakuru, R A ; Zhang, J ; Grubb, Anders LU
; Su, J
; Gentz, R
and Abrahamson, Magnus
LU
(1998)
In Journal of Biological Chemistry
273(38).
p.24797-24804
- Abstract
- A previously undescribed human member of the cystatin superfamily called cystatin F has been identified by expressed sequence tag sequencing in human cDNA libraries. A full-length cDNA clone was obtained from a library made from mRNA of CD34-depleted cord blood cells. The sequence of the cDNA contained an open reading frame encoding a putative 19-residue signal peptide and a mature protein of 126 amino acids with two disulfide bridges and enzyme-binding motifs homologous to those of Family 2 cystatins. Unlike other human cystatins, cystatin F has 2 additional Cys residues, indicating the presence of an extra disulfide bridge stabilizing the N-terminal region of the molecule. Recombinant cystatin F was produced in a baculovirus expression... (More)
- A previously undescribed human member of the cystatin superfamily called cystatin F has been identified by expressed sequence tag sequencing in human cDNA libraries. A full-length cDNA clone was obtained from a library made from mRNA of CD34-depleted cord blood cells. The sequence of the cDNA contained an open reading frame encoding a putative 19-residue signal peptide and a mature protein of 126 amino acids with two disulfide bridges and enzyme-binding motifs homologous to those of Family 2 cystatins. Unlike other human cystatins, cystatin F has 2 additional Cys residues, indicating the presence of an extra disulfide bridge stabilizing the N-terminal region of the molecule. Recombinant cystatin F was produced in a baculovirus expression system and characterized. The mature recombinant protein processed by insect cells had an N-terminal segment 7 residues longer than that of cystatin C and displayed reversible inhibition of papain and cathepsin L (Ki = 1.1 and 0.31 nM, respectively), but not cathepsin B. Like cystatin E/M, cystatin F is a glycoprotein, carrying two N-linked carbohydrate chains at positions 36 and 88. An immunoassay for quantification of cystatin F showed that blood contains low levels of the inhibitor (0.9 ng/ml). Six B cell lines in culture secreted barely detectable amounts of cystatin F, but several T cell lines and especially one myeloid cell line secreted significant amounts of the inhibitor. Northern blot analysis revealed that the cystatin F gene is primarily expressed in peripheral blood cells and spleen. Tissue expression clearly different from that of the ubiquitous inhibitor, cystatin C, was also indicated by a high incidence of cystatin F clones in cDNA libraries from dendritic and T cells, but no clones identified by expressed sequence tag sequencing in several B cell libraries and in >600 libraries from other human tissues and cells. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1113018
- author
- Ni, J
; Alvarez Fernandez, Marcia
LU
; Danielsson, Lena
; Chillakuru, R A
; Zhang, J
; Grubb, Anders
LU
; Su, J
; Gentz, R
and Abrahamson, Magnus
LU
- organization
- publishing date
- 1998
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 273
- issue
- 38
- pages
- 24797 - 24804
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:9733783
- scopus:0032544743
- ISSN
- 1083-351X
- language
- English
- LU publication?
- yes
- id
- 2e213f1b-8bce-495c-8b26-e6e900d4be65 (old id 1113018)
- alternative location
- http://www.jbc.org/cgi/content/full/273/38/24797
- date added to LUP
- 2016-04-01 12:09:26
- date last changed
- 2023-01-03 04:39:54
@article{2e213f1b-8bce-495c-8b26-e6e900d4be65,
abstract = {{A previously undescribed human member of the cystatin superfamily called cystatin F has been identified by expressed sequence tag sequencing in human cDNA libraries. A full-length cDNA clone was obtained from a library made from mRNA of CD34-depleted cord blood cells. The sequence of the cDNA contained an open reading frame encoding a putative 19-residue signal peptide and a mature protein of 126 amino acids with two disulfide bridges and enzyme-binding motifs homologous to those of Family 2 cystatins. Unlike other human cystatins, cystatin F has 2 additional Cys residues, indicating the presence of an extra disulfide bridge stabilizing the N-terminal region of the molecule. Recombinant cystatin F was produced in a baculovirus expression system and characterized. The mature recombinant protein processed by insect cells had an N-terminal segment 7 residues longer than that of cystatin C and displayed reversible inhibition of papain and cathepsin L (Ki = 1.1 and 0.31 nM, respectively), but not cathepsin B. Like cystatin E/M, cystatin F is a glycoprotein, carrying two N-linked carbohydrate chains at positions 36 and 88. An immunoassay for quantification of cystatin F showed that blood contains low levels of the inhibitor (0.9 ng/ml). Six B cell lines in culture secreted barely detectable amounts of cystatin F, but several T cell lines and especially one myeloid cell line secreted significant amounts of the inhibitor. Northern blot analysis revealed that the cystatin F gene is primarily expressed in peripheral blood cells and spleen. Tissue expression clearly different from that of the ubiquitous inhibitor, cystatin C, was also indicated by a high incidence of cystatin F clones in cDNA libraries from dendritic and T cells, but no clones identified by expressed sequence tag sequencing in several B cell libraries and in >600 libraries from other human tissues and cells.}},
author = {{Ni, J and Alvarez Fernandez, Marcia and Danielsson, Lena and Chillakuru, R A and Zhang, J and Grubb, Anders and Su, J and Gentz, R and Abrahamson, Magnus}},
issn = {{1083-351X}},
language = {{eng}},
number = {{38}},
pages = {{24797--24804}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Cystatin F is a glycosylated human low molecular weight cysteine proteinase inhibitor}},
url = {{http://www.jbc.org/cgi/content/full/273/38/24797}},
volume = {{273}},
year = {{1998}},
}