Production of N-Lauroylated G Protein .ALPHA.-Subunit in Sf9 Insect Cells: The Type of N-Acyl Group of G.ALPHA. Influences G Protein-Mediated Signal Transduction
スポンサーリンク
概要
- 論文の詳細を見る
The α-subunit of rod photoreceptor G protein transducin (Gt1α) is heterogeneously modified at the N-terminus by a mixture of acyl groups, laurate (C12:0), myristate (C14:0), and two unsaturated fatty acids (C14:1 and C14:2). Although the N-fatty acylation of Gt1α plays important roles in protein-protein and protein-membrane interactions in light signaling, the biological significance of the heterogeneous acylation remains unclear due to the difficulty in isolating each Gt1α isoform from the retinal rod cells. Here we found that Gt1α/Gi1α chimera (Gt/iα) expressed in Sf9 cells is also heterogeneously modified by myristate (_??_90%) and laurate (_??_10%), raising the possibility that the N-acyl group of recombinant Gt/iα may be manipulated by modifying culture media. In fact, addition of myristic acid to the medium decreased the relative content of lauroylated Gt/iα to an undetectable level, whereas exogenously added lauric acid significantly increased the relative content of lauroylated Gt/iα in a concentrationdependent manner. By culturing the Gt/iα-virus infected Sf9 cells with fatty acids, we obtained four different preparations of N-acylated Gt/iα, in which the relative abundance of lauroylated isoform was 0%, 20%, 33% and _??_70%, respectively. Functional analysis of these proteins showed that an increase in the relative content of the lauroylated isoform remarkably slowed down the steady-state GTP hydrolysis rate of Gt/iα; the steady-state GTPase activity of the lauroylated isoform was estimated to be one order of magnitude lower than that of the myristoylated isoform. These results suggest that the retinal Gt1α is composed of isoforms having functionally heterogeneous signaling properties.
- 社団法人 日本生化学会の論文
著者
-
Haga Tatsuya
Institute For Bimolecular Science Faculty Of Science Gakushuin University
-
Matsuura Yoshiharu
Research Center For Emerging Infectious Diseases Research Institute For Microbial Diseases Osaka Uni
-
Matsuda Takahiko
Department Of Biophysics And Biochemistry Graduate School Of Science The University Of Tokyo
-
Fukada Yoshitaka
Department Of Biophysics And Biochemistry Graduate School Of Science The University Of Tokyo
-
Hashimoto Yuichi
Department Of Biophysics And Biochemistry Graduate School Of Science The University Of Tokyo
関連論文
- 5-Oxo-Eicosatetraenoic Acid-Induced Chemotaxis : Identification of a Responsible Receptor hGPCR48 and Negative Regulation by G Protein G_
- Molecular Cloning and Characterization of a cDNA Encoding the Retinal Arylalkylamine N-Acetyltransferase of the Rainbow Trout, Oncorhynchus mykiss
- LOCALIZATION OF CRY4, A NOVEL BLUE-LIGHT PHOTORECEPTOR-LIKE PROTEIN, IN THE CHICKEN PINEAL GLAND(Physiology,Abstracts of papers presented at the 76^ Annual Meeting of the Zoological Society of Japan)
- A NOVEL PHOTORECEPTOR CANDIDATE EXPRESSED IN THE CHICKEN RETINA AND PINEAL GLAND(Physiology,Abstracts of papers presented at the 75^ Annual Meeting of the Zoological Society of Japan)
- ENZYMATIC PROPERTIES OF THE OPSIN KINASE IN THE ZEBRAFISH CONE CELLS(Physiology,Abstracts of papers presented at the 76^ Annual Meeting of the Zoological Society of Japan)
- Immunohistochemical Localization of Opsins and Alpha-Subunit of Transducin in the Pineal Complex and Deep Brain of the Japanese Grass Lizard, Takydromus tachydromoides(Physiology)
- Immunoreactivities to Rhodopsin and Rod/Cone Transducin Antisera in the Retina, Pineal Complex and Deep Brain of the Bullfrog, Rana catesbeiana
- Production of N-Lauroylated G Protein α-Subunit in Sf9 Insect Cells : The Type of N-Acyl Group of Gα Influences G Protein-Mediated Signal Transduction
- A CLOCK-RELATED GENE Lcg EXPRESSED IN THE CHICKEN PINEAL GLAND(Physiology,Abstracts of papers presented at the 76^ Annual Meeting of the Zoological Society of Japan)
- ANALYSIS OF PUTATIVE CLOCK-RELATED GENES INDUCED BY LIGHT IN A PHASE-DEPENDENT MANNER(Physiology&Biochemistry,Abstracts of papers presented at the 74^ Annual Meeting of the Zoological Society of Japan)
- Light-Dependent Changes in the Chick Pineal Temperature and the Expression of cHsp90αgene : A Potential Contribution of in vivo Temperature Change to the Photic-Entrainment of the Chick Pineal Circadian Clock(Physiology)
- REGULATION OF PHOTOTRANSDUCTION BY SELECTED FARNESYLATION OF PHOTORECEPTOR G PROTEIN γ-SUBUNIT(Physiology,Abstracts of papers presented at the 75^ Annual Meeting of the Zoological Society of Japan)
- Regulatory Mechanism for the Stability of the Meta II Intermediate of Pinopsin
- The Differences in the Expressions of Visual Pigments and Transducin in Photoreceptor Cell Differentiation
- Functional Role of Melatonin in the Formation of Circadian Rhythms in the Zebrafish(Oscillatory Mechanisms and Functional Differentiation of Circadian Clocks,Symposium,PROCEEDING OF THE 76^ ANNUAL MEETING OF THE ZOOLOGICAL SOCIETY OF JAPAN)
- Effect of Brefeldin A on Melatonin Secretion of Chick Pineal Cells^1
- Chicktacking Pineal Clock
- SCREENING FOR TRANSCRIPTION FACTORS REGULATING THE ZEBRAFISH PINEAL PHOTORECEPTOR CELL DIFFERENTIATION(Developmental Biology,Abstracts of papers presented at the 76^ Annual Meeting of the Zoological Society of Japan)
- COMPARISON OF GENE EXPRESSION PROFILES BETWEEN THE ZEBRAFISH PINEAL AND RETINAL PHOTORECEPTOR CELLS(Physiology,Abstracts of papers presented at the 74^ Annual Meeting of the Zoological Society of Japan)
- TRANSGENIC ZEBRAFISH EXPRESSING DOMINANT NEGATIVE MUTANT OF zfBMAL IN THE PINEAL GLAND(Physiology&Biochemistry,Abstracts of papers presented at the 74^ Annual Meeting of the Zoological Society of Japan)
- PIPE : A NOVEL CIS-ACTING ELEMENT RESPONSIBLE FOR PINEAL-SPECIFIC GENE EXPRESSION IN THE ZEBRAFISH(Physiology)(Proceedings of the Seventy-Third Annual Meeting of the Zoological Society of Japan)
- IN VIVO ANALYSIS OF THE ZEBRAFISH Bmal FUNCTION IN THE PINEAL CLOCK SYSTEM(Physiology)(Proceedings of the Seventy-Third Annual Meeting of the Zoological Society of Japan)
- ESTABLISHMENT OF TRANSGENIC ZEBRAFISH HAVING PIPE-INDUCING GFP EXPRESSION IN THE PINEAL GLAND(Physiology,Abstracts of papers presented at the 76^ Annual Meeting of the Zoological Society of Japan)
- Molecular Oscillation Behind the Clockface
- ROLE OF MAP-KINASE IN THE OSCILLATION MECHANISM OF THE CHICKEN PINEAL CIRCADIAN CLOCK(Physiology)(Proceedings of the Seventy-Third Annual Meeting of the Zoological Society of Japan)
- Resetting Mechanism of Central and Peripheral Circadian Clocks in Mammals
- W4-4 Chick Pineal Circadian Clock : The Clock Genes and Photoentrainment.
- Production of N-Lauroylated G Protein .ALPHA.-Subunit in Sf9 Insect Cells: The Type of N-Acyl Group of G.ALPHA. Influences G Protein-Mediated Signal Transduction