The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal.

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Abstract Text:

K Murakami,K Mihara,T Omura,

Microsomal-type cytochrome P450s are integral membrane proteins bound to the membrane through their N-terminal transmembrane hydrophobic segment, the signal anchor sequence. To elucidate the determinants that enable the P450s to be located in the ER, we constructed cDNAs encoding chimeric proteins in which a secretory form of carboxyesterase, carboxyesterase Sec, was connected to the N-terminus of the full-length or truncated forms of a microsomal-type P450, P450(M1), and the constructed plasmids were expressed in COS cells. Since carboxyesterase Sec is an N-glycosylated secretory protein, endo H treatment could be used to determine whether these chimeric proteins were located in the ER or not. Carboxyesterase Sec with the N-terminal 20 amino acids, containing the transmembrane region, of P450(M1), was located in the ER, as determined from the endo H sensitivity of the expressed protein and immunofluorescence staining of the cells. As the expressed protein exhibited carboxyesterase activity, it was not retained in the ER through the BiP-dependent quality control system recognizing unfolded proteins. Another chimeric protein construct in which carboxyesterase Sec was connected to the C-terminal region of rat UDP-glucuronosyltransferase (UDP-GT), that contained a double-lysin ER retention motif, was also located in the ER, as determined from the endo H sensitivity and immunofluorescence staining. On the other hand, the sugar moiety of the carboxyesterase Sec connected to the transmembrane segment of UDP-GT, Sec/GTd, was partially resistant to the endo H treatment. From the results of immunofluorescent staining and cell fractionation, it was concluded that the Sec/GTd product was located in the Golgi apparatus. These observations indicated that the N-terminal hydrophobic segment of P450(M1) is sufficient for the ER membrane retention, whereas the transmembrane segment of UDP-GT is not. To determine whether microsomal P450s are recycled between the ER and Golgi compartments or not, a DNA construct encoding cathepsin D connected to the N-terminus of P450(M1) was prepared and expressed in COS cells. The fusion protein was phosphorylated, but the phosphorylation was sensitive to alkaline phosphatase. As a control, authentic cathepsin D was subjected to phosphorylation of its oligosaccharide chain that was resistant to the alkaline phosphatase treatment. Since GlcNAc-P-transferase, which forms the alkaline phosphatase-resistant phosphodiester in the sugar chains of lysosome-targeting proteins, is located in the Golgi apparatus, it was concluded that the oligosaccharide chain of the cathepsin D portion of the fusion protein was not phosphorylated, and that the chimeric protein did not go to the Golgi apparatus.(ABSTRACT TRUNCATED AT 400 WORDS)

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Publishing Authors By Initials

K Murakami,K Mihara,T Omura,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

PUBMED ID PMID:

MEDLINE DATE:

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 116

Page Numbers: 164-75

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jul

YEAR: 1994

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: chemistry

Chemical & Substance for Abstract: The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. Information

Substance Name: Cathepsin D

Registry Number: EC 3.4.23.5

Grant and Affiliation Information for The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal.

AFFILIATION: Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka.

Country: JAPAN

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: J Biochem

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal Related Publications

• Acetanilide-hydrolyzing esterase of rat liver microsomes. I. Solubilization, purification, and intramicrosomal localization.
• Acetanilide-hydrolyzing esterase of rat liver microsomes. II. Turnover studies.
• Actinocatenispora sera sp. nov., isolated by long-term culturing.
• Actinohivin, a novel anti-HIV protein from an actinomycete that inhibits syncytium formation: isolation, characterization, and biological activities.
• Biogenesis of endoplasmic reticulum membrane in rat liver cells. I. Intracellular sites of synthesis of cytochrome b5 and NADPH-cytochrome c reductase.
• Biogenesis of endoplasmic reticulum membrane in rat liver cells. II. Discharge of the nascent peptides of NADPH-cytochrome c reductase and cytochrome b5 on the cytoplasmic side of the endoplasmic reticulum membrane.
• Biogenesis of the mitochondrial matrix enzyme, glutamate dehydrogenase, in rat liver cells. I. Subcellular localization, biosynthesis, and intracellular translocation of glutamate dehydrogenase.
• Biogenesis of the mitochondrial matrix enzyme, glutamate dehydrogenase, in rat liver cells. II. Significance of binding of glutamate dehydrogenase to microsomal membrane.
• Bone with a vascular flap induced from fat tissue with the use of rhBMP-2 in rats.
• Design and synthesis via click chemistry of 8,9-anhydroerythromycin A 6,9-hemiketal analogues with anti-MRSA and -VRE activity.
• Different turnover behavior of phenobarbital-induced and normal NADPH-cytochrome c reductases in rat liver microsomes.
• Distribution of hepatic sulfite oxidase among subcellular organelles and its intraorganelle localization.
• Inhibition of 6-methylsalicyclic acid synthesis by the antibiotic cerulenin.
• Inhibition of the biosynthesis of leucomycin, a macrolide antibiotic, by cerulenin.
• Isolation and Structure Elucidation of Seco-neokadsuranic Acid A and 3,4-Seco-(24Z)-lanosta-4(30),8,24-triene-3,26-dioic Acid1.
• Isolation and structure elucidation of 12beta-acetoxycoccinic Acid, 12beta-hydroxycoccinic Acid, 12alpha-acetoxycoccinic Acid, and 12alpha-hydroxycoccinic acid1.
• Microbacterium sediminicola sp. nov. and Microbacterium marinilacus sp. nov., isolated from marine environments.
• Potential therapeutics for obesity and atherosclerosis: inhibitors of neutral lipid metabolism from microorganisms.
• Presence of apo-cytochrome beta 5 in microsomes from rat liver.
• Prevalence of Primary Aldosteronism: Should We Screen for Primary Aldosteronism before Treating Hypertensive Patients with Medication?
• Purification and Chemical Properties of Anti-complementary Polysaccharide from the Leaves of Artemisia princeps.
• Purification and properties of NADH-cytochrome b5 reductase solubilized by lysosomes from rat liver microsomes.
• Solubilization of NADH-cytochrome b5 reductase from liver microsomes by lysosomal digestion.
• Stability of messenger RNA's for microsomal NADPH-cytochrome c reductase and cytochrome b 5 in the livers of normal and phenobarbital-treated rats.
• Structural Characterization of Anti-Complementary Polysaccharides from the Leaves of Artemisia princeps.
• Subfractionation of rat liver microsomes by immunoprecipitation and immunoadsorption methods.
• The early stage of labeling of microsomal membrane proteins in rat liver by radioactive amino acids.
• The nucleases from Acrocylindrium sp. I. Purification and properties of ribonuclease.
• Triterpenoid Acids from Kadsura longipedunculata. Neokadsuranic Acids B and C: Two Novel Triterpenoids with 14 (13 --> 12)abeo-Lanostane Skeletons.
• Yemuoside YM7, YM11, YM13, and YM14: Four Nortriterpenoid Saponins from Stauntonia chinensis.