全文获取类型
收费全文 | 2520篇 |
免费 | 64篇 |
国内免费 | 48篇 |
出版年
2023年 | 11篇 |
2022年 | 12篇 |
2021年 | 26篇 |
2020年 | 33篇 |
2019年 | 27篇 |
2018年 | 58篇 |
2017年 | 43篇 |
2016年 | 44篇 |
2015年 | 55篇 |
2014年 | 123篇 |
2013年 | 174篇 |
2012年 | 136篇 |
2011年 | 179篇 |
2010年 | 120篇 |
2009年 | 129篇 |
2008年 | 121篇 |
2007年 | 135篇 |
2006年 | 127篇 |
2005年 | 103篇 |
2004年 | 108篇 |
2003年 | 64篇 |
2002年 | 51篇 |
2001年 | 36篇 |
2000年 | 36篇 |
1999年 | 36篇 |
1998年 | 32篇 |
1997年 | 47篇 |
1996年 | 61篇 |
1995年 | 62篇 |
1994年 | 38篇 |
1993年 | 31篇 |
1992年 | 36篇 |
1991年 | 38篇 |
1990年 | 24篇 |
1989年 | 37篇 |
1988年 | 19篇 |
1987年 | 22篇 |
1986年 | 19篇 |
1985年 | 32篇 |
1984年 | 40篇 |
1983年 | 32篇 |
1982年 | 30篇 |
1981年 | 16篇 |
1980年 | 13篇 |
1979年 | 4篇 |
1978年 | 4篇 |
1977年 | 2篇 |
1976年 | 1篇 |
1974年 | 3篇 |
1973年 | 1篇 |
排序方式: 共有2632条查询结果,搜索用时 453 毫秒
61.
Processing of Procarboxypeptidase E into Carboxypeptidase E Occurs in Secretory Vesicles 总被引:2,自引:0,他引:2
Abstract: Carboxypeptidase E (CPE) functions in the posttranslational processing of bioactive peptides. Like other peptide processing enzymes, CPE is initially produced as a precursor ("proCPE") that undergoes posttranslational processing at a site containing five adjacent Arg residues near the N-terminus and at other sites near the C-terminus of proCPE. The time course of the N-terminal processing step suggests that this conversion occurs in either the Golgi apparatus or the secretory vesicles. To delineate further the site of proCPE processing, pulse/chase analysis was performed under conditions that block transit out of the Golgi apparatus (brefeldin A, carbonyl cyanide m -chlorophenylhydrazone, or 20°C) or that block acidification of vesicles (chloroquine, monensin, or ammonium chloride). The results of these analysis suggest that efficient proCPE processing requires an acidic post-Golgi compartment. To test whether known processing enzymes can perform this cleavage, purified proCPE was incubated with furin, prohormone convertase 1, or a dynorphin converting enzyme, and the products were analyzed on denaturing polyacrylamide gels. Furin cleaves proCPE within the N-terminal region, although the reaction is not very efficient, requiring relatively large amounts of furin or long incubation times. The other two peptide processing enzymes did not cleave proCPE, whereas a relatively small amount of secretory granule extract was able to convert proCPE into CPE. Taken together, these findings suggest that the conversion of proCPE into CPE occurs primarily in secretory vesicles. 相似文献
62.
While the insect fragment count is currently the primary test used for assessing insect contamination of food products, this
technique is very problematical for assaying microalgal materials. An account is given of a new immunoassay technique,which
is based on an enzyme-linked immunosorbent assay(ELISA) detection of insect myosin and which provides a rapid and convenient
means of quantitatively determining the amount of insect contamination in algal product samples with a high degree of replicability.
Up to 30 samples can be tested in duplicate in 2.5–3 h. Experiments were carried out with a variety of common contaminant
insects of algal products, using various life stages, including Corixidae, Ephydridaeand Chironomidae using both Spirulina
(Arthrospira) and Chlorella as typical algal materials. As little as one insect per 50 g sample can readily be detected, with
excellent correlation (r2 = 0.99) between the number of insects present and the color produced. A matrix analysis to determine the ruggedness of the
immunoassay was carried out following the protocols of the AOAC International and established that minor departures in seven
variables from the standard assay resulted in no substantial differences. The insect myosin assay offers a quantitative and
reliable means for assessing insect contamination of algal materials and should be considered for adoption as a standard method
for this type of product.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
63.
Summary To study the influence of phosphorylation and oxidation on the repeat domains of human Tau protein, we faced the challenge to selectively dimerize two cysteine-containing peptides in the presence of a nearby phosphate group. To this end, we were able to demonstrate the utility of a selective dimerization approach by forming disulfide bonds in unprotected phosphopeptides and extended the methodology to unprotected glycopeptides. Activation of one cysteine of a peptide chain with 2,2-dithiodipyridine and coupling this thiopyridyl-peptide to another peptide chain, containing an unprotected cysteine residue, yielded the mixed dimers in high purities and reasonable yields. Phosphate or sugar side chains on either peptide component remained unaffected during the activation and dimerization processes. While for mixed dimers the activated peptides were isolated by chromatography, homodimers were obtained by a simple one-pot reaction after 1 h. We demonstrate that cysteines can be dimerized in unprotected phosphopeptides and glycopeptides, without any side reactions affecting these posttranslational modifications.Abbreviations DCM
dichloromethane
- DMF
N,N-dimethylformamide
- DTP
2,2-dithiodipyridine
- Fmoc
9-fluorenylmethyloxycarbonyl
- HPLC
high-performance liquid chromatography
- MALDI
matrix-assisted laser desorption/ionization
- MS
mass spectrometry
- NFT
neurofibrillary tangles
- PHF
paired helical filaments
- PKC
protein kinase C
- RP
reversed phase
-
human Tau protein
- TFA
trifluoroacetic acid
Parts of this paper were presented at the 24th European Peptide Symposium in Edinburgh, Scotland, U.K., September 8–13, 1996. 相似文献
64.
Summary Proton chemical shifts of a series of disordered linear peptides (H-Gly-Gly-X-Gly-Gly-OH, with X being one of the 20 naturally
occurring amino acids) have been obtained using 1D and 2D 1H NMR at pH 5.0 as a function of temperature and solvent composition. The use of 2D methods has allowed some ambiguities in
side-chain assignments in previous studies to be resolved. An additional benefit of the temperature data is that they can
be used to obtain ‘random coil’ amide proton chemical shifts at any temperature between 278 and 318 K by interpolation. Changes
of chemical shift as a function of trifluoroethanol concentration have also been determined at a variety of temperatures for
a subset of peptides. Significant changes are found in backbone and side-chain amide proton chemical shifts in these ‘random
coil’ peptides with increasing amounts of trifluoroethanol, suggesting that caution is required when interpreting chemical
shift changes as a measure of helix formation in peptides in the presence of this solvent. Comparison of the proton chemical
shifts obtained here for H-Gly-Gly-X-Gly-Gly-OH with those for H-Gly-Gly-X-Ala-OH [Bundi, A. and Wüthrich, K. (1979) Biopolymers, 18, 285–297] and for Ac-Gly-Gly-X-Ala-Gly-Gly-NH2 [Wishart, D.S., Bigam, C.G., Holm, A., Hodges, R.S. and Sykes, B.D. (1995) J. Biomol. NMR, 5, 67–81] generally shows good agreement for CH protons, but reveals significant variability for NH protons. Amide proton chemical
shifts appear to be highly sensitive to local sequence variations and probably also to solution conditions. Caution must therefore
be exercised in any structural interpretation based on amide proton chemical shifts. 相似文献
65.
Michael D. Reily Venkataraman Thanabal Michael E. Adams 《Journal of biomolecular NMR》1995,5(2):122-132
Summary The 48 amino acid peptides -Aga-IVA and -Aga-IVB are the first agents known to specifically block P-type calcium channels in mammalian brain, thus complementing the existing suite of pharmacological tools used for characterizing calcium channels. These peptides provide a new set of probes for studies aimed at elucidating the structural basis underlying the subtype specificity of calcium channel antagonists. We used 288 NMR-derived constraints in a protocol combining distance geometry and molecular dynamics employing the program DGII, followed by energy minimization with Discover to derive the three-dimensional structure of -Aga-IVB. The toxin consists of a well-defined core region, comprising seven solvent-shielded residues and a well-defined triple-stranded -sheet. Four loop regions have average backbone rms deviations between 0.38 and 1.31 Å, two of which are well-defined type-II -turns. Other structural features include disordered C- and N-termini and several conserved basic amino acids that are clustered on one face of the molecule. The reported structure suggests a possible surface for interaction with the channel. This surface contains amino acids that are identical to those of another known P-type calcium channel antagonist, -Aga-IVA, and is rich in basic residues that may have a role in binding to the anionic sites in the extracellular regions of the calcium channel.Abbreviations TOCSY
total correlated spectroscopy
- NOESY
nuclear Overhauser enhancement spectroscopy
- COSY
correlated spectroscopy 相似文献
66.
Summary Nuclear Overhauser effect (NOE) measurements on molecules in solution provide information about only the ensemble-averaged properties of these molecules. An algorithm is presented that uses a list of NOEs to produce an ensemble of molecules that on average agrees with these NOEs, taking into account the effect of surrounding spins on the buildup of each NOE (spin diffusion). A simplified molecular dynamics simulation on several copies of the molecule in parallel is restrained by forces that are derived directly from differences between calculated and measured NOEs. The algorithm is tested on experimental NOE data of a helical peptide derived from bovine pancreatic trypsin inhibitor. 相似文献
67.
Izaura Yoshico Hirata Maria Helena Sedenho Cezari Clovis Ryuichi Nakaie Paulo Boschcov Amando Siuiti Ito Maria Aparecida Juliano Luiz Juliano 《Letters in Peptide Science》1995,1(6):299-308
Summary A general procedure, using the commonly employed solid-phase peptide synthesis methodology for obtaining internally quenched fluorogenic peptides with ortho-aminobenzoyl/dinitrophenyl groups as donor-acceptor pairs, is presented. The essential feature of this procedure is the synthesis of an N
-Boc or-Fmoc derivative of glutamic acid with the -carboxyl group bound to N-(2,4-dinitrophenyl)-ethylenediamine (EDDnp), which provides the quencher moiety attached to the C-terminus of the substrate. The fluorescent donor group, ortho-aminobenzoic acid (Abz), is incorporated into the resin-bound peptide in the last coupling cycle. Depending on the resin type used, Abz-peptidyl-Gln-EDDnp or Abz-peptidyl-Glu-EDDnp is obtained. Using the procedure described above, substrates for human renin and tissue kallikreins were synthesised. Spectrofluorimetric measurements of Abz bound to the -amino group of proline showed that strong quenching of Abz fluorescence occurs in the absence of any acceptor group. 相似文献
68.
Immunochemical study on PHI/PHM with use of synthetic peptides 总被引:2,自引:0,他引:2
N Yanaihara C Yanaihara K Nokihara K Iguchi S Fukata M Tanaka Y Yamamoto T Mochizuki 《Peptides》1984,5(2):247-254
We have synthesized PHI and PHM (human PHI) as well as their fragments, PHI (1-6), PHI (1-15), PHI (14-19), PHI (14-27), PHI (20-27), PHM (1-15) and PHM (13-27), by the solution or solid-phase method for peptide synthesis. Using the highly purified synthetic peptides as immunogens or haptenic immunogens, five kinds of PHI/PHM specific antisera were produced. The major antibody-recognition sites of the five antisera were located respectively in the PHI C-terminal (R8201), in the PHI N-terminal (R8403), in the PHM C-terminal (R8502), and in the PHM whole molecule (R8702 and R8703). Radioimmunoassays (RIAs) with antisera R8201, R8403 and R8502, respectively, showed a wide distribution of immunoreactive (IR) PHI/PHM in porcine and human gastrointestinal and brain tissues. The concentrations of IR-PHI in the porcine gastrointestinal tissues, however, differed between the R8201 and R8403 RIAs employed for measurement. By using these two different PHI RIAs, the IR-PHI in the porcine brain tissue extract was shown to be almost a single component coeluting with synthetic PHI in gel filtration. The IR-PHI in the extract of porcine lower intestine on the other hand, contained, besides a PHI-like component, unidentified component(s) eluting immediately after synthetic PHI in gel filtration; this crossreacted with the PHI C-terminal specific R8201 antiserum but not with the N-terminal specific R8403 antiserum, suggesting the presence of the C-terminal-related fragment(s) of PHI in the tissues. 相似文献
69.
Determination of the glucosidase-stimulating proteins by competitive enzyme-linked immunoassay 总被引:1,自引:0,他引:1
A procedure for the immunoassay of cohydrolase sphingolipid-I in mouse tissue is described. This cohydrolase (actually a mixture of at least four related proteins) stimulates or activates the beta-glucosidase which hydrolyzes ceramide glucoside, a widely occurring glycosphingolipid. The method involves extraction of cohydrolase from tissue homogenate with a salt-buffer solution, removal of proteins by adjustment to pH 6, further removal of proteins by heating, and removal of interfering materials with a small size exclusion column. Antibodies were raised to bovine cohydrolase in rabbits and purified with an affinity column made from cohydrolase. The immunoassay involves binding of antibody by the cohydrolase sample (20-200 pg) in competition with cohydrolase that has been chemically linked to horseradish peroxidase. The mixture is treated with particle-linked second antibody and centrifuged; the pellet is then assayed fluorometrically for peroxidase content. Initial application of the method showed that cohydrolase was present in all mouse tissues studied and that its concentration paralleled that of glucocerebrosidase relatively closely. Changes with age (14 and 92 days) occurred in a similar fashion for the two substances. 相似文献
70.
alpha-Endorphin and gamma-endorphin, two closely related peptides of the pro-opiomelanocortin family with characteristic biological activities, were purified to homogeneity from single human pituitary glands and chemically identified. Isolation of the peptides was based on size fractionation by Sephadex G-75 chromatography followed by two HPLC steps using reverse-phase and paired-ion reverse-phase systems and was monitored by radioimmunoassay. During the isolation procedure alpha- and gamma-endorphin-sized material behaved chromatographically and immunologically indistinguishably from synthetic alpha- and gamma-endorphin. The amino acid composition and NH2-terminus of isolated peptides demonstrated their identity as authentic alpha-endorphin and gamma-endorphin. Acetylated forms were absent. In addition, evidence is provided that large forms with alpha- and gamma-endorphin immunoreactivity detected during gel filtration are human lipotropin-(1-74) and -(1-75), respectively. The data substantiate that alpha-endorphin and gamma-endorphin exist as endogenous peptides in the human pituitary gland. 相似文献