Correlation between leucine rich domain and the stability of LRWD1 protein in human NT2/D1 cells

Yung Chieh Tsai, Yen Ni Teng, Jui Hsiang Hung, Chien Hsing Wu, Yu Ting Kuo, Pao Lin Kuo, Chien Chih Chiu, Bin Huang

Research output: Contribution to journalArticle

Abstract

Purpose: LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. Materials and methods: The NT2/D1 cells separately transfected with full length of LRWD1 gene (LRWDWT) or genes with deleted sequences in the LRR domain (LRWD1ΔLRR), WD1 domain (LRWD1ΔWD1), WD2 domain (LRWD1ΔWD2), WD3 domain (LRWD1ΔWD3) and entire three WD domains (LRWD1Δ3xWD) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. Results: Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. Conclusions: The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.

Original languageEnglish
Pages (from-to)266-272
Number of pages7
JournalAdvances in Medical Sciences
Volume59
Issue number2
DOIs
Publication statusPublished - 2014 Sep 1

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Leucine
Proteins
Receptors, Tumor Necrosis Factor, Type II
human LRWD1 protein
Protein Stability
Proteasome Endopeptidase Complex
Spermatogenesis
Immunoprecipitation
Genes
Mass Spectrometry
Flow Cytometry
Tumor Necrosis Factor-alpha
Western Blotting

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Tsai, Yung Chieh ; Teng, Yen Ni ; Hung, Jui Hsiang ; Wu, Chien Hsing ; Kuo, Yu Ting ; Kuo, Pao Lin ; Chiu, Chien Chih ; Huang, Bin. / Correlation between leucine rich domain and the stability of LRWD1 protein in human NT2/D1 cells. In: Advances in Medical Sciences. 2014 ; Vol. 59, No. 2. pp. 266-272.
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abstract = "Purpose: LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. Materials and methods: The NT2/D1 cells separately transfected with full length of LRWD1 gene (LRWDWT) or genes with deleted sequences in the LRR domain (LRWD1ΔLRR), WD1 domain (LRWD1ΔWD1), WD2 domain (LRWD1ΔWD2), WD3 domain (LRWD1ΔWD3) and entire three WD domains (LRWD1Δ3xWD) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. Results: Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. Conclusions: The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.",
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Correlation between leucine rich domain and the stability of LRWD1 protein in human NT2/D1 cells. / Tsai, Yung Chieh; Teng, Yen Ni; Hung, Jui Hsiang; Wu, Chien Hsing; Kuo, Yu Ting; Kuo, Pao Lin; Chiu, Chien Chih; Huang, Bin.

In: Advances in Medical Sciences, Vol. 59, No. 2, 01.09.2014, p. 266-272.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Correlation between leucine rich domain and the stability of LRWD1 protein in human NT2/D1 cells

AU - Tsai, Yung Chieh

AU - Teng, Yen Ni

AU - Hung, Jui Hsiang

AU - Wu, Chien Hsing

AU - Kuo, Yu Ting

AU - Kuo, Pao Lin

AU - Chiu, Chien Chih

AU - Huang, Bin

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Y1 - 2014/9/1

N2 - Purpose: LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. Materials and methods: The NT2/D1 cells separately transfected with full length of LRWD1 gene (LRWDWT) or genes with deleted sequences in the LRR domain (LRWD1ΔLRR), WD1 domain (LRWD1ΔWD1), WD2 domain (LRWD1ΔWD2), WD3 domain (LRWD1ΔWD3) and entire three WD domains (LRWD1Δ3xWD) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. Results: Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. Conclusions: The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.

AB - Purpose: LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. Materials and methods: The NT2/D1 cells separately transfected with full length of LRWD1 gene (LRWDWT) or genes with deleted sequences in the LRR domain (LRWD1ΔLRR), WD1 domain (LRWD1ΔWD1), WD2 domain (LRWD1ΔWD2), WD3 domain (LRWD1ΔWD3) and entire three WD domains (LRWD1Δ3xWD) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. Results: Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. Conclusions: The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.

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