Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)

碩士 === 國立中興大學 === 生命科學系所 === 97 === The peptidylarginine deiminase (PAD) is a Ca2+-dependent enzyme that can carry out protein citrullination (deimination) which is the conversion of protein-bound arginine to citrulline (Arg -> Cit). PAD4 is the only one isotype with a nuclear localization signal...

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Main Authors: Yu-Hsiu Chiang, 江玉秀
Other Authors: 洪慧芝
Format: Others
Language:en_US
Published: 2009
Online Access:http://ndltd.ncl.edu.tw/handle/96013070947453584192
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spelling ndltd-TW-097NCHU51050062016-04-29T04:19:42Z http://ndltd.ncl.edu.tw/handle/96013070947453584192 Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4) 探討人類第四型胜肽精胺酸去亞胺酶的四級結構組成與酵素催化之間關係 Yu-Hsiu Chiang 江玉秀 碩士 國立中興大學 生命科學系所 97 The peptidylarginine deiminase (PAD) is a Ca2+-dependent enzyme that can carry out protein citrullination (deimination) which is the conversion of protein-bound arginine to citrulline (Arg -> Cit). PAD4 is the only one isotype with a nuclear localization signal (NLS) region and the gene of PAD4 has a susceptible locus for rheumatoid arthritis (RA). The structures of PAD4 combined without or with calcium ions and the substrate have been resolved by X-ray crystallography. PAD4 is a dimer consisted of two monomer head-to-tail contact. However, it is still unclear the relationship between enzyme regulation and subunit-subunit interaction of PAD4. To gain insight into the quaternary structure stability of PAD4, we used LIGPLOT and DIMPLOT to find the important residues at dimer interface of PAD4 with hydrogen-bonded interaction. The Arg8 of one monomer is hydrogen-bonded with Asp547 of the other monomer and these two amino acid residues are charged residues containing the ionic interaction. On the other hand, the Tyr435 of one PAD4 monomer also has the hydrogen-bonded interaction with Tyr237 and Glu281 on the other monomer. Then we analyzed a series of PAD4 mutations at residues Arg8, Tyr237, Glu281, Tyr435, and Asp547 on the dimer interface by site-directed mutagenesis to disrupt the dimer organization combined with the analytical ultracentrifugation data and kinetic analysis. The mutations of PAD4 dimer interface caused quaternary structural change accompanied by the decreased activities and Hill coefficient (h) compared to WT. Here, our data indicated that the role of the dimeric state of PAD4 is not only important to maintain the stability of the overall structure but also to regulate its catalysis. Moreover, the ionic interaction between Arg8 and Asp547 slightly affect the dimer interface and the hydrophobic interaction of Tyr435 is the most important factor for the structure stability and activity of PAD4. 洪慧芝 2009 學位論文 ; thesis 58 en_US
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description 碩士 === 國立中興大學 === 生命科學系所 === 97 === The peptidylarginine deiminase (PAD) is a Ca2+-dependent enzyme that can carry out protein citrullination (deimination) which is the conversion of protein-bound arginine to citrulline (Arg -> Cit). PAD4 is the only one isotype with a nuclear localization signal (NLS) region and the gene of PAD4 has a susceptible locus for rheumatoid arthritis (RA). The structures of PAD4 combined without or with calcium ions and the substrate have been resolved by X-ray crystallography. PAD4 is a dimer consisted of two monomer head-to-tail contact. However, it is still unclear the relationship between enzyme regulation and subunit-subunit interaction of PAD4. To gain insight into the quaternary structure stability of PAD4, we used LIGPLOT and DIMPLOT to find the important residues at dimer interface of PAD4 with hydrogen-bonded interaction. The Arg8 of one monomer is hydrogen-bonded with Asp547 of the other monomer and these two amino acid residues are charged residues containing the ionic interaction. On the other hand, the Tyr435 of one PAD4 monomer also has the hydrogen-bonded interaction with Tyr237 and Glu281 on the other monomer. Then we analyzed a series of PAD4 mutations at residues Arg8, Tyr237, Glu281, Tyr435, and Asp547 on the dimer interface by site-directed mutagenesis to disrupt the dimer organization combined with the analytical ultracentrifugation data and kinetic analysis. The mutations of PAD4 dimer interface caused quaternary structural change accompanied by the decreased activities and Hill coefficient (h) compared to WT. Here, our data indicated that the role of the dimeric state of PAD4 is not only important to maintain the stability of the overall structure but also to regulate its catalysis. Moreover, the ionic interaction between Arg8 and Asp547 slightly affect the dimer interface and the hydrophobic interaction of Tyr435 is the most important factor for the structure stability and activity of PAD4.
author2 洪慧芝
author_facet 洪慧芝
Yu-Hsiu Chiang
江玉秀
author Yu-Hsiu Chiang
江玉秀
spellingShingle Yu-Hsiu Chiang
江玉秀
Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
author_sort Yu-Hsiu Chiang
title Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
title_short Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
title_full Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
title_fullStr Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
title_full_unstemmed Study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (PAD4)
title_sort study on the relationship between the quaternary structure organizations and enzyme catalysis of human peptidylarginine deiminase 4 (pad4)
publishDate 2009
url http://ndltd.ncl.edu.tw/handle/96013070947453584192
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