TY - JOUR
T1 - Stability and Splitting Produces Revealed by Matrix-Assisted Laser Desorption Ionization-Time of Flight-Mass Spectrometry in Human Serum Transferrin
AU - Zhuo, Hui Qin
AU - Jin, Hong Wei
AU - Huang, He Qing
AU - Huang, Hui Ying
AU - CAI, Zongwei
N1 - Funding Information:
Received 25 October 2006; accepted 31 January 2007 * Corresponding author. Email: [email protected]; Tel: +86 592-2184614 This work was supported by the National Natural Science Foundation of China (No. 30470372), the Foundation of Xiamen University of China (Nos. 2004xdcx207 and 2005xdkjcx 20051009), and the Foundation of Key Laboratory of MOD for Cell Biology and Tumor Cell Engineering, Xiamen University of China (No. 2005-113). Copyright © 2007, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences. Published by Elsevier Limited. All rights reserved.
PY - 2007/6
Y1 - 2007/6
N2 - Human serum transferrin (HTF) has been prepared to study its molecular weight by analysis of mass spectrometry. Several analytical techniques, such as SDS-PAGE, in-solution enzymatic digestion, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), database search, and comparison approaches were used to identify stability and cleavage products amongst iron-binding HTF (IB-HTF), such as iron-saturated HTF (IS-HTF), HTF-2Fe(III), and apoHTF. By using acetonitrile as the eluant, IS-HTF and HTF-2Fe(III) showed cleavage behavior, although separated by reversed-phase HPLC (RP-HPLC). IS-HTF and HTF-2Fe(III) were decomposed into various cleavage products of peptides comprising different molecular weights while treated with acetonitrile, indicating that the structural stability in HTF is tightly connected with the iron numbers binding the protein. The iron can improve the stability of the molecular structure in HTF. By comparison, the regulation and pathway of forming various peptides from the HTF were studied for revealing the cleavage mechanism while being treated with acetonitrile. Under the same treating condition, the approach of protein refolding gave rise to various HTF polymers and its cleavage products. It is suggested that these polymers and products interfere with diagnosing both congenital disorders of glycosylation and chronic alcohol abuse accurately in the clinic.
AB - Human serum transferrin (HTF) has been prepared to study its molecular weight by analysis of mass spectrometry. Several analytical techniques, such as SDS-PAGE, in-solution enzymatic digestion, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), database search, and comparison approaches were used to identify stability and cleavage products amongst iron-binding HTF (IB-HTF), such as iron-saturated HTF (IS-HTF), HTF-2Fe(III), and apoHTF. By using acetonitrile as the eluant, IS-HTF and HTF-2Fe(III) showed cleavage behavior, although separated by reversed-phase HPLC (RP-HPLC). IS-HTF and HTF-2Fe(III) were decomposed into various cleavage products of peptides comprising different molecular weights while treated with acetonitrile, indicating that the structural stability in HTF is tightly connected with the iron numbers binding the protein. The iron can improve the stability of the molecular structure in HTF. By comparison, the regulation and pathway of forming various peptides from the HTF were studied for revealing the cleavage mechanism while being treated with acetonitrile. Under the same treating condition, the approach of protein refolding gave rise to various HTF polymers and its cleavage products. It is suggested that these polymers and products interfere with diagnosing both congenital disorders of glycosylation and chronic alcohol abuse accurately in the clinic.
KW - Biochemical marker
KW - Human serum transferrin
KW - In-solution enzymatic digestion
KW - Matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS)
KW - Reversed phase high performance liquid chromatography
UR - http://www.scopus.com/inward/record.url?scp=34447259305&partnerID=8YFLogxK
U2 - 10.1016/S1872-2040(07)60057-7
DO - 10.1016/S1872-2040(07)60057-7
M3 - Journal article
AN - SCOPUS:34447259305
SN - 1872-2040
VL - 35
SP - 791
EP - 796
JO - Chinese Journal of Analytical Chemistry
JF - Chinese Journal of Analytical Chemistry
IS - 6
ER -