scholarly journals Cloning, Expression, and Functional Characterization of a Ca2+-dependent Endoplasmic Reticulum Nucleoside Diphosphatase

2002 ◽  
Vol 277 (40) ◽  
pp. 36978-36986 ◽  
Author(s):  
Bernd U. Failer ◽  
Norbert Braun ◽  
Herbert Zimmermann
Keyword(s):  
Endoplasmic Reticulum ◽  
Functional Characterization ◽  
Nucleoside Diphosphatase

Identification and functional characterization of a novel human protein highly related to the yeast dynamin-like GTPase Vps1p

1998 ◽  
Vol 111 (10) ◽  
pp. 1341-1349 ◽  
Author(s):  
M. Imoto ◽  
I. Tachibana ◽  
R. Urrutia
Keyword(s):  
Endoplasmic Reticulum ◽  
Cho Cells ◽  
Mammalian Cells ◽  
Functional Characterization ◽  
Luciferase Reporter ◽  
Pleckstrin Homology ◽  
Rich Region ◽  
Gtpase Domain ◽  
Proline Rich Region

Dynamin proteins containing a GTPase domain, a pleckstrin homology motif and a proline-rich tail participate in receptor-mediated endocytosis in organisms ranging from insects to vertebrates. In addition, dynamin-related GTPases, such as the yeast Golgi protein Vps1p, which lack both the pleckstrin homology motif and the proline-rich region, participate in vesicular transport within the secretory pathway in lower eukaryotes. However, no data is available on the existence of Vps1p-like proteins in mammalian cells. In this study, we report the identification and characterization of a novel gene encoding a human dynamin-related protein, DRP1, displaying high similarity to the Golgi dynamin-like protein Vps1p from yeast and to a Caenorhabditis elegans protein deposited in the databank. These proteins are highly conserved in their N-terminal tripartite GTPase domain but lack the pleckstrin homology motif and proline-rich region. Northern blot analysis reveals that the DRP1 mRNA is detected at high levels in human muscle, heart, kidney and brain. Immunolocalization studies in Chinese hamster ovary (CHO) cells using an epitope-tagged form of DRP1 and confocal microscopy show that this protein is concentrated in a perinuclear region that labels with the endoplasmic reticulum marker DiOC6(3) and the Golgi marker C5-DMB-Cer. In addition, the localization of DRP1 is highly similar to the localization of the endoplasmic reticulum and cis-Golgi GTPase Rab1A, but not to the staining for the trans-Golgi GTPase Rab6. Furthermore, overexpression of a cDNA encoding a GTP binding site mutant of DRP1 (DRP1(K38E)) in CHO cells decreases the amount of a secreted luciferase reporter protein, whereas the overexpression of wild-type DRP1 increases the secretion of this marker. Together, these results constitute the first structural and functional characterization of a mammalian protein similar to the yeast dynamin-related GTPase Vps1p and indicate that the participation of these proteins in secretion has been conserved throughout evolution.

scholarly journals Functional Characterization of ERp18, a New Endoplasmic Reticulum-located Thioredoxin Superfamily Member

2003 ◽  
Vol 278 (31) ◽  
pp. 28912-28920 ◽  
Author(s):  
Heli I. Alanen ◽  
Richard A. Williamson ◽  
Mark J. Howard ◽  
Anna-Kaisa Lappi ◽  
Heli P. Jäntti ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Functional Characterization

Functional characterization of orchardgrass endoplasmic reticulum-resident Hsp90 (DgHsp90) as a chaperone and an ATPase

2009 ◽  
Vol 47 (10) ◽  
pp. 859-866 ◽  
Author(s):  
Joon-Yung Cha ◽  
Min Hee Jung ◽  
Netty Ermawati ◽  
Mukhamad Su'udi ◽  
Gyu-Jin Rho ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Functional Characterization

scholarly journals Structural and functional characterization of Sec66p, a new subunit of the polypeptide translocation apparatus in the yeast endoplasmic reticulum.

1993 ◽  
Vol 4 (9) ◽  
pp. 931-939 ◽  
Author(s):  
D Feldheim ◽  
K Yoshimura ◽  
A Admon ◽  
R Schekman
Keyword(s):  
Endoplasmic Reticulum ◽  
Signal Sequence ◽  
Functional Characterization ◽  
Yeast Cells ◽  
Temperature Sensitive ◽  
Endoplasmic Reticulum Membrane ◽  
Restrictive Temperature ◽  
Permissive Temperature

SEC66 encodes the 31.5-kDa glycoprotein of the Sec63p complex, an integral endoplasmic reticulum membrane protein complex required for translocation of presecretory proteins in Saccharomyces cerevisiae. DNA sequence analysis of SEC66 predicts a 23-kDa protein with no obvious NH2-terminal signal sequence but with one domain of sufficient length and hydrophobicity to span a lipid bilayer. Antibodies directed against a recombinant form of Sec66p were used to confirm the membrane location of Sec66p and that Sec66p is a glycoprotein of 31.5 kDa. A null mutation in SEC66 renders yeast cells temperature sensitive for growth. sec66 cells accumulate some secretory precursors at a permissive temperature and a variety of precursors at the restrictive temperature. sec66 cells show defects in Sec63p complex formation. Because sec66 cells affect the translocation of some, but not all secretory precursor polypeptides, the role of Sec66p may be to interact with the signal peptide of presecretory proteins.

Isolation and functional characterization of Sporothrix schenckii ROT2, the encoding gene for the endoplasmic reticulum glucosidase II

2012 ◽  
Vol 116 (8) ◽  
pp. 910-918 ◽  
Author(s):  
Claudia I. Robledo-Ortiz ◽  
Arturo Flores-Carreón ◽  
Arturo Hernández-Cervantes ◽  
Aurelio Álvarez-Vargas ◽  
Keunsook K. Lee ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Functional Characterization ◽  
Sporothrix Schenckii ◽  
Glucosidase Ii ◽  
Encoding Gene

scholarly journals Functional Characterization of Arabidopsis Calreticulin1a: A Key Alleviator of Endoplasmic Reticulum Stress

2008 ◽  
Vol 49 (6) ◽  
pp. 912-924 ◽  
Author(s):  
A. Christensen ◽  
K. Svensson ◽  
S. Persson ◽  
J. Jung ◽  
M. Michalak ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Endoplasmic Reticulum Stress ◽  
Functional Characterization

Cloning and characterization of the heart muscle isoform of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) from crayfish

2002 ◽  
Vol 205 (17) ◽  
pp. 2677-2686 ◽  
Author(s):  
Dongdong Chen ◽  
Zhiping Zhang ◽  
Michele G. Wheatly ◽  
Yongping Gao
Keyword(s):  
Amino Acids ◽  
Endoplasmic Reticulum ◽  
Heart Muscle ◽  
Transmembrane Domain ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Northern Analysis ◽  
Specific Probe ◽  
Tail Muscle

SUMMARY This paper describes the cloning and functional characterization of the heart muscle isoform of Sarco/endoplasmic reticulum Ca2+-ATPase(SERCA) from crayfish Procambarus clarkii. The complete crayfish heart SERCA, identified by reverse transcription-polymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends (RACE), consists of 4495 bp with a 3060 bp open reading frame, coding for 1020 amino acids. This isoform differs from the previously identified axial abdominal (tail) muscle SERCA solely in its C-terminal amino acids. The last nine amino acids of the tail muscle isoform are replaced by 27 hydrophobic amino acids in the heart isoform that have the potential to form an additional transmembrane domain. Consistent with other invertebrate studies, Southern blot analysis suggested that the heart and tail muscle isoforms are encoded from the same gene that is equally related to SERCA-1, -2 and -3 of vertebrates. The tissue distributions of these two isoforms have been assessed using isoform-specific probes and northern analysis. A cardiac-specific probe bound only to a 5.8 kb species in heart and had minimal cross-hybridization with 7.6 and 5.8 kb species in eggs and no hybridization with tail muscle. A tail-isoform-specific probe hybridized with a 4.5 kb species in tail muscle and cross-hybridized with a 4.5 kb species in eggs and 8.8 kb in heart muscle. Both isoforms are expressed in eggs suggesting that transcripts are formed early in development and are subsequently broadly expressed in all tissue types. Expression of the cardiac muscle SERCA isoform varied with the stage of moulting. Expression was high in intermoult and decreased in premoult. However, expression was restored rapidly in postmoult (within 2 days) unlike expression of tail muscle SERCA,which remained downregulated for weeks. Differences in contractility between the two muscle types in the postmoult period may explain these expression patterns.

scholarly journals Functional characterization of alternatively spliced human SERCA3 transcripts

1998 ◽  
Vol 275 (6) ◽  
pp. C1449-C1458 ◽  
Author(s):  
Esteban Poch ◽  
Stephen Leach ◽  
Susan Snape ◽  
Tasha Cacic ◽  
David H. MacLennan ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibody ◽  
Endoplasmic Reticulum ◽  
Amino Acid ◽  
Functional Characterization ◽  
Lymphoid Tissues ◽  
Gene Products ◽  
Alternatively Spliced ◽  
Lower Capacity

The sarcoplasmic (or endoplasmic) reticulum Ca2+-ATPase (SERCA)-3 has been implicated in the possible dysregulation of Ca2+ homeostasis that accompanies the pathology of hypertension and diabetes. We report the molecular cloning of two alternatively spliced transcripts from the human SERCA3 gene, ATP2A3, that encode proteins that differ at their carboxy termini by 36 amino acids. SERCA3 transcripts were most abundantly expressed in lymphoid tissues, intestine, pancreas, and prostate. The two human SERCA3 proteins encoded by alternatively spliced transcripts were recognized by the monoclonal antibody PL/IM430 and demonstrated Ca2+ uptake and ATPase activity with an apparent Ca2+ affinity 0.5 pCa unit lower than that of other SERCA gene products. The subcellular distribution of SERCA3 protein was indistinguishable from that of SERCA2b, with expression in the nuclear envelope and in the endoplasmic reticulum throughout the cell. Two variant SERCA3 constructs, huS3-I and huS3-II, were isolated that encode proteins with three amino acid differences: Ala-673 (in huS3-I) substituted for Thr (in huS3-II), Ile-817 substituted for Met, and an insertion of Glu-994. huS3-I displayed a 10-fold lower capacity to transport Ca2+ than huS3-II.

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