지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제6호
- 발행연도
- 2015.1
- 수록면
- 782 - 787 (6page)
이용수
초록· 키워드
오류제보하기
In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah- cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.
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참고문헌 (23)
참고문헌 신청
Arya R / 2008 / Dictyostelium discoideum - a promising expression system for the production of eukaryotic proteins / FASEB J 22 : 4055 ~ 4066
Bjørgo E / 1998 / Partial characterization and three-dimensional structural localization of eight mutations in exon 7 of the human phenylalanine hydroxylase gene associated
Choi YK / 2005 / D-Threo tetrahydrobiopterin is synthesized via 1’-oxo-2’-D-hydroxytetrahydropterin in Dictyostelium discoideum Ax2 / FEBS Lett 579 : 3085 ~ 3089
Erlandsen H / 2003 / Structural studies on phenylalanine hydroxylase and implications toward understanding and treating phenylketonuria / Pediatrics 112 : 1557 ~ 1565
Fitzpatrick PF / 2012 / Allosteric regulation of phenylalanine hydroxylase / Arch. Biochem. Biophys 519 : 194 ~ 201
Bjørgo E / 1998 / Partial characterization and three-dimensional structural localization of eight mutations in exon 7 of the human phenylalanine hydroxylase gene associated
Choi YK / 2005 / D-Threo tetrahydrobiopterin is synthesized via 1’-oxo-2’-D-hydroxytetrahydropterin in Dictyostelium discoideum Ax2 / FEBS Lett 579 : 3085 ~ 3089
Erlandsen H / 2003 / Structural studies on phenylalanine hydroxylase and implications toward understanding and treating phenylketonuria / Pediatrics 112 : 1557 ~ 1565
Fitzpatrick PF / 2012 / Allosteric regulation of phenylalanine hydroxylase / Arch. Biochem. Biophys 519 : 194 ~ 201
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