کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10557267 | 968501 | 2005 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Application of matrix-assisted laser desorption and ionization time-of-flight mass spectrometry for the characterization of the substrate specificity of neutrophil phospholipase A2
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کلمات کلیدی
ESIN-formyl-l-methionyl-l-leucyl-l-phenylalaninefMLPLPLPLA2PMNLPAHBSSlysophosphatidylethanolamineLPELPCTFALPSFFAPMA - LDC هاphospholipase A2 - آنزیم فسفولیپاز A2 Trifluoroacetic acid - اسید Trifluoroaceticphosphatidic acid - اسید فسفاتیدیکlysophosphatidic acid - اسید لیسفسفیدیدFree fatty acid - اسید چرب آزادfast-atom bombardment - بمباران سریع اتمSubstrate specificity - خصوصیات زیربناییdihydroxybenzoic acid - دی هیدروکسی بنزوئیک اسیدFab - فابphosphatidylcholine - فسفاتیدیل کولینphosphatidylethanolamine - فسفاتیدیلتانولامینPhosphatidylserine - فسفاتیدیلسرینPhospholipids - فسفولیپیدPhospholipid - فسفولیپیدpolymorphonuclear leukocytes - لکوسیت های پلی مورفونیک هسته ایLysophosphatidylcholine - لیزوفسفاتیدیل کولینLysophosphatidylserine - لیسفسفاتیدیلسرینlysophospholipid - لیسفسفلیپیدMALDI-TOF MS - مالدی توف MSHank's balanced salt solution - محلول نمک متعادل هانکneutrophil - نوتروفیلelectrospray ionisation - یونیزاسیون الکترو اسپری
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) was successfully applied for the analysis of various lipid classes. It can also be used for monitoring the digestion of phosphatidylcholine (PC) with phospholipase A2 (PLA2) and it was shown that MALDI-TOF MS possesses a number of advantages over well established methods for this purpose. In this work, we use MALDI-TOF MS for determination of the substrate specificity of neutrophil PLA2. For the comparison of the selectivity of the enzyme to various phospholipid (PL) classes, the intensities of the signals arising from the product of the reaction (Sp) and the signal intensity of the residual substrate (Ss) were compared and the resulting Sp/Ss ratio was used as the measure. This approach was first tested with a model system pancreatic PLA2 and afterwards two sources of the neutrophil PLA2-the enzyme extracted from the neutrophils and the enzyme released from these cells-were tested for their substrate specificity. We will show that the neutrophil-secreted PLA2 possesses high preferences for digestion of phosphatidic acid (PA) over other phospholipids. The method applied here is simple and much information can be obtained from a single mass spectrum. Moreover, this approach works well also with a crude biological systems, i.e. no prior purification of the enzyme is required for means of characterisation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Microchemical Journal - Volume 80, Issue 1, April 2005, Pages 31-37
Journal: Microchemical Journal - Volume 80, Issue 1, April 2005, Pages 31-37
نویسندگان
Marijana PetkoviÄ, Julia Müller, Jürgen Schiller, Jürgen Arnhold,