Abstract
Protein delivery to primary cells by protein transduction domain (PTD) serves as a novel measure for manipulation of the cells for biological study and for the treatment of various human conditions. Although the method has been employed to modulate cellular function in vitro, only limited reports are available on its application in the replacement of deficient signaling molecules into primary cells. We examined the potential of recombinant proteins to compensate for defective cytosolic components of the NADPH oxidase complex in chronic granulomatous disease (CGD) neutrophils in both p47 phox and p67 phox deficiency. The p47 phox or p67 phox protein linked to Hph-1 PTD was effectively expressed in soluble form and transduced into human neutrophils efficiently without eliciting unwanted signal transduction or apoptosis. The delivered protein was stable for more than 24h, expressed in the cytoplasm, translocated to the membrane fraction upon activation, and, most importantly able to restored reactive oxygen species (ROS) production. Although research on human primary neutrophils using the protein delivery system is still limited, our data show that the protein transduction approach for neutrophils may be applicable to the control of local infections in CGD patients by direct delivery of the protein product.
| Original language | English |
|---|---|
| Pages (from-to) | 162-168 |
| Number of pages | 7 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 417 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2012 Jan 6 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
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Transducible form of p47 phox and p67 phox compensate for defective NADPH oxidase activity in neutrophils of patients with chronic granulomatous disease . / Honda, Fumiko; Hane, Yumiko; Toma, Tomoko; Yachie, Akihiro; Kim, Eun Sung; Lee, Sang Kyou; Takagi, Masatoshi; Mizutani, Shuki; Morio, Tomohiro.
In: Biochemical and Biophysical Research Communications, Vol. 417, No. 1, 06.01.2012, p. 162-168.Research output: Contribution to journal › Article
TY - JOUR
T1 - Transducible form of p47 phox and p67 phox compensate for defective NADPH oxidase activity in neutrophils of patients with chronic granulomatous disease
AU - Honda, Fumiko
AU - Hane, Yumiko
AU - Toma, Tomoko
AU - Yachie, Akihiro
AU - Kim, Eun Sung
AU - Lee, Sang Kyou
AU - Takagi, Masatoshi
AU - Mizutani, Shuki
AU - Morio, Tomohiro
PY - 2012/1/6
Y1 - 2012/1/6
N2 - Protein delivery to primary cells by protein transduction domain (PTD) serves as a novel measure for manipulation of the cells for biological study and for the treatment of various human conditions. Although the method has been employed to modulate cellular function in vitro, only limited reports are available on its application in the replacement of deficient signaling molecules into primary cells. We examined the potential of recombinant proteins to compensate for defective cytosolic components of the NADPH oxidase complex in chronic granulomatous disease (CGD) neutrophils in both p47 phox and p67 phox deficiency. The p47 phox or p67 phox protein linked to Hph-1 PTD was effectively expressed in soluble form and transduced into human neutrophils efficiently without eliciting unwanted signal transduction or apoptosis. The delivered protein was stable for more than 24h, expressed in the cytoplasm, translocated to the membrane fraction upon activation, and, most importantly able to restored reactive oxygen species (ROS) production. Although research on human primary neutrophils using the protein delivery system is still limited, our data show that the protein transduction approach for neutrophils may be applicable to the control of local infections in CGD patients by direct delivery of the protein product.
AB - Protein delivery to primary cells by protein transduction domain (PTD) serves as a novel measure for manipulation of the cells for biological study and for the treatment of various human conditions. Although the method has been employed to modulate cellular function in vitro, only limited reports are available on its application in the replacement of deficient signaling molecules into primary cells. We examined the potential of recombinant proteins to compensate for defective cytosolic components of the NADPH oxidase complex in chronic granulomatous disease (CGD) neutrophils in both p47 phox and p67 phox deficiency. The p47 phox or p67 phox protein linked to Hph-1 PTD was effectively expressed in soluble form and transduced into human neutrophils efficiently without eliciting unwanted signal transduction or apoptosis. The delivered protein was stable for more than 24h, expressed in the cytoplasm, translocated to the membrane fraction upon activation, and, most importantly able to restored reactive oxygen species (ROS) production. Although research on human primary neutrophils using the protein delivery system is still limited, our data show that the protein transduction approach for neutrophils may be applicable to the control of local infections in CGD patients by direct delivery of the protein product.
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UR - http://www.scopus.com/inward/citedby.url?scp=84855758073&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2011.11.077
DO - 10.1016/j.bbrc.2011.11.077
M3 - Article
VL - 417
SP - 162
EP - 168
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -