皮摩尔水平的氨基酸分析。在多肽C末端序列分析中的应用。
Amino acid analysis at the picomole level. Application to the C-terminal sequence analysis of polypeptides.
作者信息
Chang J Y, Knecht R, Braun D G
出版信息
Biochem J. 1981 Dec 1;199(3):547-55. doi: 10.1042/bj1990547.
Abstract
Amino acids labelled with dimethylaminoazobenzenesulphonyl chloride can be separated by reversed-phase high-pressure liquid chromatography and detected in the visible region (436 nm). All 19 naturally occurring amino acids can be separated on a Zorbax ODS column by employing two different gradient systems consisting of an acetonitrile/aqueous buffer mixture. As little as 2--5 pmol of an individual dimethylaminoazobenzenesulphonyl-amino acid can be quantitatively analysed with reliability, and only 10--30 ng of the dimethylaminoazobenzenesulphonylated protein hydrolysate is needed for each complete amino acid analysis. This new technique is as sensitive as any of the current amino acid analysis methods involving ion-exchange separation plus fluorescence detection, and is technically much simpler. By the combination of this sensitive amino acid-analysing technique with carboxypeptidase, we have been able to determine the C-terminal sequence of polypeptides at the picomole level.
摘要
用二甲基氨基偶氮苯磺酰氯标记的氨基酸可通过反相高压液相色谱法分离，并在可见光区域（436纳米）进行检测。通过使用由乙腈/水性缓冲液混合物组成的两种不同梯度系统，所有19种天然存在的氨基酸都可以在Zorbax ODS柱上分离。低至2 - 5皮摩尔的单个二甲基氨基偶氮苯磺酰基氨基酸能够可靠地进行定量分析，每次完整的氨基酸分析仅需要10 - 30纳克的二甲基氨基偶氮苯磺酰化蛋白质水解产物。这项新技术与任何当前涉及离子交换分离加荧光检测的氨基酸分析方法一样灵敏，并且在技术上要简单得多。通过将这种灵敏的氨基酸分析技术与羧肽酶相结合，我们已经能够在皮摩尔水平上确定多肽的C末端序列。
相似文献
1
Amino acid analysis at the picomole level. Application to the C-terminal sequence analysis of polypeptides.
Biochem J. 1981 Dec 1;199(3):547-55. doi: 10.1042/bj1990547.
2
Isolation and characterization of polypeptide at the picomole level. Pre-column formation of peptide derivatives with dimethylaminoazobenzene isothiocyanate.
Biochem J. 1981 Dec 1;199(3):537-45. doi: 10.1042/bj1990537.
3
A complete separation of dimethylaminoazobenzenesulphonyl-amino acids. Amino acid analysis with low nanogram amounts of polypeptide with dimethylaminoazobenzenesulphonyl chloride.
Biochem J. 1982 Jun 1;203(3):803-6. doi: 10.1042/bj2030803.
4
N-terminal sequence analysis of polypeptide at the picomole level.
Biochem J. 1981 Dec 1;199(3):557-64. doi: 10.1042/bj1990557.
5
Reversed-phase high-performance liquid chromatography separation of dimethylaminoazobenzene sulfonyl- and dimethylaminoazobenzene thiohydantoin-amino acid derivatives for amino acid analysis and microsequencing studies at the picomole level.
Anal Biochem. 1989 Apr;178(1):107-17. doi: 10.1016/0003-2697(89)90364-3.
6
Analysis of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acids at sub-picomole levels by high-performance liquid chromatography: simultaneous manual sequencing of picomole quantities of several polypeptides.
Anal Biochem. 1989 Feb 15;177(1):46-9. doi: 10.1016/0003-2697(89)90011-0.
7
Analysis of physiological amino acids using dabsyl derivatization and reversed-phase liquid chromatography.
J Chromatogr. 1993 Mar 5;613(1):137-44. doi: 10.1016/0378-4347(93)80207-k.
8
Amino acid analysis in the picomole range by precolumn derivatization and high-performance liquid chromatography.
Methods Enzymol. 1983;91:41-8. doi: 10.1016/s0076-6879(83)91009-1.
9
Determination of the unusual amino acid hypusine at the lower picomole level by derivatization with 4-dimethylaminoazobenzene-4'-sulphonyl chloride and reversed-phase high-performance or medium-pressure liquid chromatography.
J Chromatogr. 1992 Jul 31;606(1):43-8. doi: 10.1016/0021-9673(92)85255-r.
10
Liquid chromatographic analysis of amino acids: using dimethylamino-azobenzenesulfonyl chloride and Hypersil ODS column to analyze the composition of apo B peptides.
J Protein Chem. 1993 Feb;12(1):11-4. doi: 10.1007/BF01024908.
引用本文的文献
1
Molecular Engineering of Bacterioferritin: A Versatile Nanodimensional Protein Cage.
Molecules. 2023 Jun 9;28(12):4663. doi: 10.3390/molecules28124663.
2
Peptidoglycan-type analysis of the N-acetylmuramic acid auxotrophic oral pathogen Tannerella forsythia and reclassification of the peptidoglycan-type of Porphyromonas gingivalis.
BMC Microbiol. 2019 Sep 2;19(1):200. doi: 10.1186/s12866-019-1575-7.
3
The influence of seed apparency, nutrient content and chemical defenses on dietary preference in Dipodomys ordii.
Oecologia. 1990 Mar;82(3):333-341. doi: 10.1007/BF00317480.
4
Possibility of production of amino acids by impact reaction using a light-gas gun as a simulation of asteroid impacts.
Orig Life Evol Biosph. 2015 Jun;45(1-2):195-205. doi: 10.1007/s11084-015-9419-4. Epub 2015 Mar 22.
5
Amino acid supplementation enhances urokinase production by HT-1080 cells.
J Ind Microbiol Biotechnol. 2014 Jun;41(6):1035-8. doi: 10.1007/s10295-014-1434-3. Epub 2014 Mar 27.
6
Differences in amino acid sequence of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from two species of Dasycladaceae.
Planta. 1986 Dec;169(4):575-82. doi: 10.1007/BF00392109.
7
Analysis of amino acids as DABS-derivatives with a sensitivity to the femtomole level using RP-HPLC narrow-bore columns.
Amino Acids. 1992 Oct;3(3):303-9. doi: 10.1007/BF00806005.
8
Growth Rate-Dependent Modulation of Carbon Flux through Central Metabolism and the Kinetic Consequences for Glucose-Limited Chemostat Cultures of Corynebacterium glutamicum.
Appl Environ Microbiol. 1996 Feb;62(2):429-36. doi: 10.1128/aem.62.2.429-436.1996.
9
Hyperhomocystinemia impairs endothelial function and eNOS activity via PKC activation.
Arterioscler Thromb Vasc Biol. 2005 Dec;25(12):2515-21. doi: 10.1161/01.ATV.0000189559.87328.e4. Epub 2005 Oct 6.
10
Effect of charge and molecular weight on transdermal peptide delivery by iontophoresis.
Pharm Res. 2005 Dec;22(12):2069-78. doi: 10.1007/s11095-005-8110-2. Epub 2005 Sep 26.
本文引用的文献
1
Procedures for the chromatographic determination of amino acids on four per cent cross-linked sulfonated polystyrene resins.
J Biol Chem. 1954 Dec;211(2):893-906.
2
A protein sequenator.
Eur J Biochem. 1967 Mar;1(1):80-91. doi: 10.1007/978-3-662-25813-2_14.
3
O-phthalaldehyde: fluorogenic detection of primary amines in the picomole range. Comparison with fluorescamine and ninhydrin.
Proc Natl Acad Sci U S A. 1975 Feb;72(2):619-22. doi: 10.1073/pnas.72.2.619.
4
Carboxypeptidase Y in sequence determination of peptides.
Methods Enzymol. 1977;47:84-93. doi: 10.1016/0076-6879(77)47010-1.
5
Subnanomole-range amino acid analysis.
Methods Enzymol. 1977;47:3-18. doi: 10.1016/0076-6879(77)47003-4.
6
Chromophoric labeling of amino acids with 4-dimethylaminoazobenzene-4'-sulfonyl chloride.
Anal Chem. 1975 Aug;47(9):1634-8. doi: 10.1021/ac60359a007.
7
Direct microsequence analysis of polypeptides using an improved sequenator, a nonprotein carrier (polybrene), and high pressure liquid chromatography.
Biochemistry. 1978 May 30;17(11):2124-33. doi: 10.1021/bi00604a016.
8
High performance liquid chromatographic determination of amino acids in the picomole range.
Anal Chem. 1979 Jul;51(8):1338-41. doi: 10.1021/ac50044a055.
9
Fluorometric microbore amino acid analyzer: the construction of an inexpensive, highly sensitive instrument using o-phthalaldehyde as a detection agent.
Anal Biochem. 1979 Jul 15;96(2):298-307. doi: 10.1016/0003-2697(79)90585-2.
10
Derivatization of cysteine and cystine for fluorescence amino acid analysis with the o-phthaldialdehyde/2-mercaptoethanol reagent.
J Biol Chem. 1979 Jul 25;254(14):6248-51.
Zotero 插件