共查询到20条相似文献,搜索用时 15 毫秒
1.
J L Omdahl K Wilson H Swerdlow W J Driscoll 《Archives of biochemistry and biophysics》1992,293(2):213-218
Porcine renodoxon is a kidney mitochondrial iron-sulfur protein (ISP) that functions to transfer electron to cytochromes P450 of the vitamin D pathway. A full-length cDNA clone to porcine renodoxin was isolated in the current investigation and used to study the protein's primary structure and immunological properties. The cysteine ligands for the iron-sulfur center, and the surface protein-binding and phosphorylation sites occupied identical positions in both porcine renodoxin and bovine adrenodoxin. Furthermore, porcine renodoxin was functionally indistinguishable from bovine adrenodoxin and the mature forms of both proteins had the same encoded length and shared approximately 91% sequence similarity. A synthetic peptide to the surface protein-binding region was used to demonstrate the antigenicity of the domain in both the porcine and the bovine ISPs. However, porcine renodoxin displayed only limited immunological identity to other regions of bovine adrenodoxin as measured by competitive enzyme-linked immunosorbent assay. Part of this immunological distinction was attributed to the COOH-terminal processing of porcine renodoxin, an action which negated expression of a COOH-terminal antigenic site that is present in bovine adrenodoxin. Other antigenic differences were linked to charged-residue substitutions that were located in predicted surface domains. The highest frequency of surface-residue substitutions in ferredoxin proteins was predicted for porcine renodoxin, which could provide a basis for understanding why the pig protein appears more antigenically divergent than other ferredoxins. 相似文献
2.
Takhar Kasumov Serguey Ilchenko Ling Li Nadia Rachdaoui Rovshan G. Sadygov Belinda Willard Arthur J. McCullough Stephen Previs 《Analytical biochemistry》2011,(1):73
We recently developed a method for estimating protein dynamics in vivo with heavy water (2H2O) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF MS) [16], and we confirmed that 2H labeling of many hepatic free amino acids rapidly equilibrated with body water. Although this is a reliable method, it required modest sample purification and necessitated the determination of tissue-specific amino acid labeling. Another approach for quantifying protein kinetics is to measure the 2H enrichments of body water (precursor) and protein-bound amino acid or proteolytic peptide (product) and to estimate how many copies of deuterium are incorporated into a product. In the current study, we used nanospray linear trap Fourier transform ion cyclotron resonance mass spectrometry (LTQ FT–ICR MS) to simultaneously measure the isotopic enrichment of peptides and protein-bound amino acids. A mathematical algorithm was developed to aid the data processing. The most notable improvement centers on the fact that the precursor/product labeling ratio can be obtained by measuring the labeling of water and a protein (or peptide) of interest, thereby minimizing the need to measure the amino acid labeling. As a proof of principle, we demonstrate that this approach can detect the effect of nutritional status on albumin synthesis in rats given 2H2O. 相似文献
3.
Marteinsson VT Reysenbach AL Birrien JL Prieur D 《Extremophiles : life under extreme conditions》1999,3(4):277-282
The whole-cell protein inventory of the deep-sea barophilic hyperthermophile Thermococcus barophilus was examined by one-dimensional SDS gradient gel electrophoresis when grown under different pressure conditions at 85°C (T
opt). One protein (P60) with a molecular mass of approximately 60 kDa was prominent at low pressures (0.3 MPa hydrostatic pressure
and 0.1 MPa atmospheric pressure) but not at deep-sea pressures (10, 30, and 40 MPa). About 17 amino acids were sequenced
from the N-terminal end of the protein. Sequence homology analysis in the GenBank database showed that P60 most closely resembled
heat-shock proteins in some sulfur-metabolizing Archaea. A high degree of amino acid identity (81%–93%) to thermosome subunits
in Thermococcales strains was found. Another protein (P35) with molecular mass of approximately 35.5 kDa was induced at 40 MPa hydrostatic
pressure but not under low-pressure conditions. No amino acid sequence homology was found for this protein when the 40 amino
acids from the N-terminal end were compared with homologous regions of proteins from databases. A PTk diagram was generated for T. barophilus. The results suggest that P
habitat is about 35 MPa, which corresponds to the in situ pressure where the strain was obtained.
Received: May 14, 1999 / Accepted: July 30, 1999 相似文献
4.
Alfred Hansel Angela Schmid Monier H. Tadros Uwe J. Jürgens 《Archives of microbiology》1994,161(2):163-167
Pore-forming protein (porin) was isolated from N,N-dimethyl-dodecylaminoxid (LDAO)-extracted outer membranes of Synechococcus PCC 6301 and purified by ion exchange chromatography on DEAE-Sephacel column. The apparent molecular mass on SDS-PAGE was determined to be about 52000. The native porin was reconstituted into black lipid bilayer membranes and showed a single-channel conductance of 5.5 nS in 1 M KCl. The porin was found to be N-terminally blocked. The C-terminal amino acid sequence was identified as Phe-Thr-Phe. Amino acid analysis suggested that the porin protein consists of about 420 amino acid residues, yielding a polarity of 43.6% and a molecular mass of 45000 in contrast to the mobility on SDS-PAGE.Abbreviations DEAE
Diethylaminoethyl; M
r, relative molecular mass
- LDAO
N,N-Dimethyl-dodecylaminoxid
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoretogram
- PCC
Pasteur Culture Collection
- SDS
sodium dodecyl sulfate
- UTEX
Culture Collection of Algae at the University of Texas 相似文献
5.
A cDNA clone encoding a 15.501 Da photosystem I (PSI) subunit of barley was isolated using an oligonucleotide based on the NH2-terminal amino acid sequence of the isolated protein. The polypeptide, which migrates with an apparent molecular mass of 9.5 kDa on denaturing SDS-PAGE, has been designated PSI-N, and the corresponding gene is PsaN. Analysis of the deduced protein sequence indicates a mature protein of 85 amino acid residues and a molecular mass of 9818 Da. PSI-N is a hydrophilic, extrinsic protein with no predicted membrane-spanning regions. The transit peptide of 60 residues (5683 Da) contains a predicted hydrophobic -helix, suggesting that the protein is routed into the thylakoid lumen. Thus, PSI-N is the second known lumenal protein component associated with PSI, together with PSI-F. 相似文献
6.
The blood clam,Barbatia lima, from Kochi, Japan, expresses a tetrameric (α 2 β 2) and a polymeric hemoglobin in erythrocytes. The latter hemoglobin is composed of unusual 34-kDa hemoglobin with a two-domain structure, and its molecular mass (about 430 kDa) is exceptionally large for an intracellular hemoglobin. The 3′ and 5′ parts of the cDNA ofB. lima two-domain globin have been amplified separately by polymerase chain reaction and the complete nucleotide sequence of 1147 bp was determined. The open reading frame is 930 nucleotides in length and encodes a protein with 309 amino acid residues, of which 73 amino acids were identified directly by protein sequencing. The mature protein begins with the acetylated Ser, and thus the N-terminus Met is cleaved. The molecular mass for the protein was calculated to be 35,244 Da. The cDNA-derived amino acid sequence ofB. lima two-domain globin shows 89% homology with that of two-domain globin fromB. reeveana, a North American species. The sequence homology between the two domains is 75%, suggesting that the two-domain globin resulted from the gene duplication of an ancestral 17-kDa globin. 相似文献
7.
The ethoxy chains of short ethoxy chain nonylphenol (NPEOav2.0, containing average 2.0 ethoxy units) were dehydrogenated by cell-free extracts from Ensifer sp. strain AS08 grown on a basal medium supplemented with NPEOav2.0. The reaction was coupled with the reduction in 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide and phenazine
methosulfate. The enzyme (NPEOav2.0 dehydrogenase; NPEO-DH) was purified to homogeneity with a yield of 20% and a 56-fold increase in specific activity. The
molecular mass of the native enzyme was 120 kDa, consisting of two identical monomer units (60 kDa). The gene encoding NPEO-DH
was cloned, which consisted of 1,659 bp, corresponding to a protein of 553 amino acid residues. The deduced amino acid sequence
agreed with the N-terminal amino acid sequence of the purified NPEO-DH. The presence of a flavin adenine dinucleotide (FAD)-binding
motif and glucose–methanol–choline (GMC) oxidoreductase signature motifs strongly suggested that the enzyme belongs to the
GMC oxidoreductase family. The protein exhibited homology (40–45% identity) with several polyethylene glycol dehydrogenases
(PEG-DHs) of this family, but the identity was lower than those (approximately 58%) among known PEG-DHs. The substrate-binding
domain was more hydrophobic compared with those of glucose oxidase and PEG-DHs. The recombinant protein had the same molecular
mass as the purified NPEO-DH and dehydrogenated PEG400-2000, NPEOav2.0 and its components, and NPEOav10, but only slight or no activity was found using diethylene glycol, triethylene glycol, and
PEG200.
English edition: The paper was edited by a native speaker through American Journal Experts (). 相似文献
8.
Hydroxyurea does not prevent synchronized G1 chinese hamster cells from entering the DNA synthetic period. 总被引:2,自引:0,他引:2
R A Walters R A Tobey C E Hildebrand 《Biochemical and biophysical research communications》1976,69(1):212-217
The amino acid sequence of subunit I of the sweet protein monellin has been completed by gas chromatographic mass spectrometry. A partial acid hydrolyzate was converted to the corresponding mixture of Nω-trifluoroethyl-polyamino alcohol-trimethylsilyl ethers and a total of 55 di- to hexapeptides were identified which could be assembled to a polypeptide chain of 44 amino acids. 相似文献
9.
Heavy chain disease proteins (FOR) were isolated from human plasma. These proteins were also detected immunochemically in
the urine of the patient. The proteins were disulphide-linked Fc-like dimers with molar mass 64.2 kg/mol and sedimentation
rate S
20,w
0
= = 0.356 ps (3.56 S). Similar amounts of aspartic acid and pyroglutamic acid were found at the N-terminus. After cyanogen
bromide cleavage of the FOR proteins, three peptides were isolated and their amino acid composition and partial amino acid
sequence was determined. We suggest that two Fc-like proteins of similar sizes are present in the plasma: (1) the first with
N-terminal aspartic acid corresponding to position 221 of γ1 EU chain and (2) the second with N-terminal pyroglutamic acid.
The first protein and small amounts of related low-molar mass fragments found also in the plasma could be degradation products
of the second protein. Evidence is given on structural differences between the FOR proteins and the corresponding portion
of the γ1 EU chain. 相似文献
10.
Tetsunori Seki 《Immunogenetics》1989,30(1):5-12
Two varieties of similar, but structurally distinct, cDNA clones for the human low-affinity receptors for the Fc portion of
immunoglobulin G (FcγRII) have been isolated. One type of clone was obtained from human B lymphocytes, and the other from
PHA-activated peripheral T cells and monocytes. Transfection of both prototype clones into Cos-7 cells and subsequent specific
staining with monoclonal antibodies of the CDw32 group confirmed the identification of the gene products. The nucleotide sequence
of the cDNA clone from B lymphocytes contains an open reading frame that encodes a protein of relative mass (M
r) 27000 with an extracellular domain of 179 amino acids containing three potential N-glycosylation sites, a 26 amino acid
transmembrane domain, and a 44 amino acid cytoplasmic domain. The clones from peripheral T cells and monocytes both encoded
a protein ofM
r 31000 with a 179 amino acid extracellular domain containing two potential N-glycosylation sites and a 26 amino acid transmembrane
domain. The two types of clones had similar sequences in their immunoglobulin-like extracellular and transmembrane domains,
but differed in their leader sequences and 3′-untranslated regions. The most notable difference between the clones was the
presence of a distinctive 76 amino acid cytoplasmic domain in those isolated from T cells and monocytes. 相似文献
11.
T Kobayashi T Takagi K Konishi K Ohnishi Y Watanabe 《European journal of biochemistry》1988,174(4):579-584
The amino acid sequence of a calcium-binding protein obtained from the cilium and cell body of Tetrahymena, designated as TCBP-10 (Tetrahymena calcium-binding protein; molecular mass = 10 kDa [Ohnishi, K. and Watanabe, Y. (1983) J. Biol. Chem. 258, 13978-13985] was determined. It is composed of 102 amino acid residues. The exact molecular mass is calculated to be 11563 Da. From the amino acid sequence analysis, it has two EF-hand-type calcium-binding sites. 相似文献
12.
13.
《FEBS letters》1986,206(1):33-35
The complete amino acid sequence of rice storage protein glutelin was determined by the sequencing of the corresponding cDNA. The deduced glutelin precursor has a 37 amino acid signal peptide sequence at the NH2 terminus, which is followed by a 269 amino acid acidic subunit (Mr = 32 489) and a 193 amino acid basic subunit (Mr = 19 587). The glutelin precursor sequence is homologous to those of pea legumin and soybean glycinin. 相似文献
14.
15.
16.
The agaA gene encoding β-agarase-a (AgaA) was cloned from the chromosomal DNA of a marine bacterium, Vibrio sp. strain PO-303. The nucleotide sequence of the agaA gene consists of 2,958 bp and encodes a protein of 985 amino acids with a molecular mass of 106,062 Da. The deduced enzyme
protein contains a typical N-terminal signal peptide of 29 amino acid residues, followed by a 266 amino acid sequence that
is homologous to catalytic module of family 16 glycoside hydrolases, a bacterial immunoglobulin group 2 (Big-2)-like domain
of 52 amino acid residues, two carbohydrate-binding modules of family 6 separated from Big-2-like domain by nine times repeated
GDDTDP amino acid sequence. AgaA is the first agarase that was identified to possess a Big-2-like domain. The recombinant
AgaA (rAgaA) expressed in Escherichia coli exhibited maximal activity around 40°C and pH 7.5, with a specific activity of 16.4 units mg−1, a K
m of 1.10 mg ml−1, and a V
max of 22.5 μmol min−1 mg−1 for agarose. The rAgaA hydrolyzed neoagarohexaose, but did not act on neoagarotetraose and neoagarobiose. 相似文献
17.
Summary The release of intact CU(I)8-thionein from copper-resistant copper-loaded yeast cells, strain X 2180-1Aa, has been shown. This copper(I)-thiolate-rich protein was characterized and compared with the chemical and physicochemical properties of intracellular yeast Cu-thionein. The same molecular mass and stoichiometry of 8 mol copper atoms/mol protein was found. No detectable difference between the Cu-thioneins was seen in luminescence emission, electronic absorption in the ultraviolet region, chiroptical data or amino acid composition. The importance of stable Cu(I)-thiolates in Cu-thionein as a safe vehicle for transporting copper in a non-reactive manner is confirmed. 相似文献
18.
Spermine binding protein (SBP) is a rat ventral prostate protein that binds various polyamines, and the level of this protein and its mRNA is regulated by androgens. Previously, the cDNA for SBP was cloned and sequenced and an amino acid sequence deduced from the cDNA. Data from cloned and sequenced and an amino acid sequence deduced from the cDNA. Data from partial amino acid sequencing of the purified protein were consistent with the amino acid sequence deduced from the cDNA. However, the amino terminus of the protein was blocked, and therefore, direct protein sequence information confirming the cDNA reading frame of this region could not be obtained by Edman degradation. We have now employed an integrated approach using fast atom bombardment mass spectrometry, tandem mass spectrometry, and conventional sequencing methodologies to establish the amino-terminal sequence of the protein and to identify an amino acid sequence (35 residues) present in the purified protein but missing from the amino acid sequence deduced from cDNA clones for this protein. The missing piece of cDNA corresponds to an exon found in mouse genomic clones for a protein similar to rat SBP. Therefore, the cDNA clones for rat SBP may represent splicing variants that lack the sequence information of one exon. The blocked amino terminus of the protein was identified as 5-oxopyrrolidine-2-carboxylic acid. Mass spectrometry also provided evidence regarding glycosylation of the protein. The first of two potential glycosylation sites clearly carries carbohydrate; the second site is, at most, only partially glycosylated. 相似文献
19.
Complete amino acid sequence of the FK506 and rapamycin binding protein, FKBP, isolated from calf thymus 总被引:19,自引:0,他引:19
FKBP, an 11.8 kD intracellular protein that binds the immunosuppressants FK506 (K
d=0.4 nM) and rapamycin (K
d=0.2 nM) with high affinity, was purified to homogeneity from calf thymus. The complete amino acid sequence has been determined by automated Edman degradation of the intact molecule and overlapping fragments generated by proteolytic and chemical cleavage. The analysis revealed a 107 amino acid peptide chain with the following sequence: GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPFKFVLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPNATLIFDVELLKLE. The molecular weight, calculated from the amino sequence to be 11,778 D, was confirmed by electrospray ionization mass spectrometry. Thus, naturally isolated bovine FKBP does not appear to have any residues modified by glycosylation, phosphorylation, or other post-translational derivatization processes. Bovine FKBP has only three amino acid residues that differ from human FKBP, whose sequence was elucidated by cloning and sequencing complementary DNA (Standaertet al., 1990). The protein has a substantial number of hydrophilic peptide segments with prevalent -strand type of chain fold. Understanding the biological function of FKBP and other members of the immunophilin class and their respective complexes with immunosuppressive drugs may provide insights into cytoplasmic signalling mechanisms, protein folding and translocation, and other cellular processes. 相似文献
20.
Ribulose 1,5-diphosphate carboxylase (RUDPcase) from autotrophically grown Euglena gracilis was purified to homogeneity as measured by analytical ultracentrifugation, polyacrylamide gel electrophoresis, and immunoprecipitation reactions. The enzyme represented about 9% of total protein and 24% of soluble protein in the autotrophic cell. Light-grown, heterotrophic cells seemed to contain considerably less RUDPcase. Native carboxylase from autotrophic Euglena showed an s20, w at low protein concentrations of 17 to 17.5, suggesting a molecular weight of >500,000 daltons. Upon denaturation, the enzyme dissociated into two subunits having different amino acid compositions and molecular weights of 59,000 and 12,000 daltons. Based upon the amino acid mass ratios, a quaternary organization of 7 to 8 large and 8 to 10 small subunits per native enzyme molecule was indicated. 相似文献