全文获取类型
收费全文 | 747176篇 |
免费 | 80899篇 |
国内免费 | 351篇 |
出版年
2018年 | 6347篇 |
2016年 | 8366篇 |
2015年 | 11149篇 |
2014年 | 13074篇 |
2013年 | 19705篇 |
2012年 | 21531篇 |
2011年 | 22112篇 |
2010年 | 14955篇 |
2009年 | 13853篇 |
2008年 | 19884篇 |
2007年 | 20562篇 |
2006年 | 19570篇 |
2005年 | 18600篇 |
2004年 | 18647篇 |
2003年 | 18003篇 |
2002年 | 17636篇 |
2001年 | 31813篇 |
2000年 | 31925篇 |
1999年 | 25849篇 |
1998年 | 9307篇 |
1997年 | 9861篇 |
1996年 | 9440篇 |
1995年 | 8705篇 |
1994年 | 8732篇 |
1993年 | 8629篇 |
1992年 | 22330篇 |
1991年 | 21702篇 |
1990年 | 21510篇 |
1989年 | 21439篇 |
1988年 | 19834篇 |
1987年 | 18821篇 |
1986年 | 17539篇 |
1985年 | 17994篇 |
1984年 | 14837篇 |
1983年 | 12838篇 |
1982年 | 9805篇 |
1981年 | 8767篇 |
1980年 | 8359篇 |
1979年 | 14231篇 |
1978年 | 11090篇 |
1977年 | 10262篇 |
1976年 | 9845篇 |
1975年 | 10709篇 |
1974年 | 11478篇 |
1973年 | 11181篇 |
1972年 | 10375篇 |
1971年 | 9312篇 |
1970年 | 8111篇 |
1969年 | 7799篇 |
1968年 | 7186篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
941.
R A Deems D Lombardo B P Morgan E D Mihelich E A Dennis 《Biochimica et biophysica acta》1987,917(2):258-268
Manoalide, a natural product from sponge, displays anti-inflammatory activity in vivo. Previous work has shown that manoalide is also a potent covalent inhibitor of the extracellular phospholipase A2 from cobra venom and that the inhibition correlated with a pH-dependent change in manoalide (Lombardo and Dennis (1985) J. Biol. Chem. 260, 7234-7240). Manoalide contains two rings and the opening of either would produce an alpha,beta-unsaturated aldehyde. The cobra venom phospholipase A2 may be able to catalyze the opening or isomerization of one of these rings, raising the possibility that manoalide is acting as a suicide substrate. To ascertain the role of the gamma-lactone ring in the inhibition, we have now investigated a synthetic manoalide analogue, 3(cis,cis-7,10)-hexadecadienyl-4-hydroxy-2-butenolide (HDHB) which contains only the alpha,beta-unsaturated gamma-lactone ring. We have found that the closed and open forms are in rapid equilibrium between pH 4 and 9 with the cyclic form being preferred at acidic pH values and the open cis form preferred at pH 9.5. When the pH is raised above 12, the alpha,beta double bond isomerizes to form trans-HDHB. Once the trans compound is formed, it is stable at all pH values and does not recyclize to the gamma-lactone ring. The observed pKa of 7.7 found for the inhibition of manoalide agrees well with the transition of the closed to the cis form of the gamma-lactone ring. Kinetic experiments with the HDHB compound show that under conditions in which the cis and closed form of the inhibitor are present in equal molar ratios, HDHB is not an irreversible inhibitor, but reversibly competes with substrate. However, the kinetics of this inhibition are complex and do not follow either pure competitive or non-competitive inhibition. The trans-HDHB exhibits similar complex kinetic but is several times more potent. The distinct differences between the behavior of manoalide and HDHB clearly indicate that while the gamma-lactone ring may play an important role in manoalide inhibition, it alone does not produce irreversible inhibition. 相似文献
942.
Subunit composition and ATP site labeling of the coated vesicle proton-translocating adenosinetriphosphatase 总被引:3,自引:0,他引:3
The partially purified proton-translocating adenosinetriphosphatase [(H+)-ATPase] from clathrin-coated vesicles has been reported to contain eight polypeptides of molecular weights 15,000-116,000 [Xie, X.S., & Stone, D.K. (1986) J. Biol. Chem. 261, 2492-2495]. To determine whether these polypeptides form a single macromolecular complex, we have isolated three monoclonal antibodies which recognize the reconstitutively active (H+)-ATPase in the native, detergent-solubilized state. All three monoclonal antibodies precipitate the same set of polypeptides from either the partially purified enzyme or the detergent-solubilized coated vesicle membrane proteins. The immunoprecipitated polypeptides have molecular weights of 100,000, 73,000, 58,000, 40,000, 38,000, 34,000, 33,000, 19,000, and 17,000. These results thus indicate that this set of polypeptides forms a single macromolecular complex and suggest that they correspond to subunits of the coated vesicle (H+)-ATPase. To identify the ATP-hydrolytic subunit of the coated vesicle (H+)-ATPase, the purified enzyme was reacted with N-ethylmaleimide (NEM) and 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl), both of which inhibit activity in an ATP-protectable manner. Labeling was carried out by using [3H]NEM or [14C]NBD-Cl, and the specificity of the reaction was increased by prelabeling of the protein with the nonradioactive reagents in the presence of ATP and by taking advantage of the nucleotide specificity of protection. The principal polypeptide labeled by both [3H]NEM and [14C]NBD-Cl had a molecular weight of 73,000. In addition, this protein was the only polypeptide whose labeling was significantly reduced in the presence of ATP.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
943.
944.
945.
946.
947.
948.
949.
950.
In an effort to facilitate studies of the reaction involved in the removal of fatty acids from acyl proteins, we have synthesized an octanoic acid ester of doubly blocked serine, specifically octanoyl N-carbobenzoxy-L-serine-benzyl ester (octanoyl boc-serine), and used it as a substrate to guide the purification of an esterase from rat lung. The esterase was purified 228-fold by column chromatography on DE-52 cellulose, hydroxylapatite, octyl-Sepharose, and concanavalin A-Sepharose and by HPLC gel filtration. The final enzyme preparation ran as a single 77,000-Da band when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and exhibited a single symmetrical peak (sedimentation coefficient, 4.5 S) when centrifuged through a sucrose density gradient (empirical Mr, 63,000). The esterase is an acidic protein, pI 4.1, and is very active against p-nitrophenyl esters comprised of C4-C14 fatty acids; the highest specific activity (26.5 mumol/min/mg) was obtained using p-nitrophenyl caprylate as substrate. The pH optimum of the lung esterase is near 8.0 and the activity on octanoyl boc-serine is maximum when 0.3% (w/v) Myrj-52 is included in the assay medium. The activity of the esterase is not dependent on calcium ions. The enzyme does not remove acyl groups from the G-protein of vesicular stomatitis virus or the proteolipid of bovine brain. The possible role of the esterase in the metabolism of acylated proteins is considered. 相似文献