Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis

Deng, Zhenzhen; Dong, Mingming; Wang, Yan; Dong, Jing; Li, Shawn S.‐C.; Zou, Hanfa; Ye, Mingliang

doi:10.1021/acs.analchem.6b04288

Cited by 15 publications

(13 citation statements)

References 22 publications

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“…[15], indicating the importance of a combined strategy. The SH2 super-binder 972 enrichment strategy has been combined with IMAC with Ti 4+ -IMAC [103], and an updated one-step SH2 super-binder method [105], as well as in a biphasic affinity chromatography approach, where Src SH2 super-binder was coupled with NeutrAvidin affinity chromatography, which resulted in enhanced specific selectivity [106].…”

Section: Imac or Moac With Antibody-based Treatmentmentioning

confidence: 99%

Phosphopeptide enrichment for phosphoproteomic analysis - A tutorial and review of novel materials

Qiu

Evans

Landels

et al. 2020

Analytica Chimica Acta

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Post-translational modifications (PTMs), mass spectrometry (MS), liquid chromatography (LC), tandem MS (MS/MS), phosphopeptide (p-peptide), label-free quantification (LFQ), stable isotope labelling by amino acids in cell culture (SILAC), Tandem Mass Tags (TMT) isobaric tags for relative and absolute quantification (iTRAQ), phospho-serine (pSer), threonine (pThr), tyrosine (pTyr), immobilized metal ion affinity chromatography (IMAC), metal oxide affinity chromatography (MOAC), acetonitrile (ACN), sodium dodecyl sulfate -polyacrylamide gel electrophoresis (SDS-PAGE), filter assisted sample preparation (FASP), solid phase extraction (SPE), high performance liquid chromatography (HPLC), reverse phase (RP), strong cation exchange (SCX), electrostatic repulsion hydrophilic interaction chromatography (ERLIC), solution isoelectric focusing (sIEF), strong anion exchange (SAX), histidine (pHis), arginine (Arg),

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Section: Imac or Moac With Antibody-based Treatmentmentioning

confidence: 99%

Phosphopeptide enrichment for phosphoproteomic analysis - A tutorial and review of novel materials

Qiu

Evans

Landels

et al. 2020

Analytica Chimica Acta

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“…This method takes a long time to balance the column. [3]. To address these aforementioned disadvantages, temperature-responsive chromatography has been developed.…”

Section: Introductionmentioning

confidence: 99%

Combination of Three Functionalized Temperature-Sensitive Chromatographic Materials for Serum Protein Analysis

Sun

Dai

et al. 2019

Molecules

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We have developed a methodology to capture acidic proteins, alkaline proteins, and glycoproteins separately in mouse serum using a combination of three functionalized temperature-responsive chromatographic stationary phases. The temperature-responsive polymer poly(N-isopropylacrylamide) was attached to the stationary phase, silica. The three temperature-responsive chromatographic stationary phase materials were prepared by reversible addition–fragmentation chain transfer polymerization. Alkaline, acidic, and boric acid functional groups were introduced to capture acidic proteins, alkaline proteins, and glycoproteins, respectively. The protein enrichment and release properties of the materials were examined using the acidic protein, bovine serum albumin; the alkaline protein, protamine; and the glycoprotein, horseradish peroxidase. Finally, the three materials were used to analyze mouse serum. Without switching the mobile phase, the capture and separation of mouse serum was achieved by the combination of three temperature-responsive chromatographic stationary phase materials. On the whole, 313 proteins were identified successfully. The number of different proteins identified using the new method was 1.46 times greater than the number of proteins that has been identified without applying this method. To our knowledge, this method is the first combinatorial use of three functionalized temperature-responsive chromatographic stationary phase silica materials to separate proteins in mouse serum.

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“…Conventional drugs are generally directed against a single gene or protein and most of melanoma patients develop drug resistance eventually [ 13 ]. SH2 superbinder, strongly binding with diverse pY sites to block related signal transduction pathways, can recognize other sites instead, even if mutation alters the pY site [ 14 , 15 ]. Therefore, SH2 superbinder can achieve the purpose of killing cancer cells [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

(Arg)9-SH2 superbinder: a novel promising anticancer therapy to melanoma by blocking phosphotyrosine signaling

Liu

Gao

et al. 2018

J Exp Clin Cancer Res

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BackgroundMelanoma is a malignant tumor with high misdiagnosis rate and poor prognosis. The bio-targeted therapy is a prevailing method in the treatment of melanoma; however, the accompanying drug resistance is inevitable. SH2 superbinder, a triple-mutant of the Src Homology 2 (SH2) domain, shows potent antitumor ability by replacing natural SH2-containing proteins and blocking multiple pY-based signaling pathways. Polyarginine (Arg)9, a powerful vector for intracellular delivery of large molecules, could transport therapeutic agents across cell membrane. The purpose of this study is to construct (Arg)9-SH2 superbinder and investigate its effects on melanoma cells, expecting to provide potential new approaches for anti-cancer therapy and overcoming the unavoidable drug resistance of single-targeted antitumor agents.Methods(Arg)9 and SH2 superbinder were fused to form (Arg)9-SH2 superbinder via genetic engineering. Pull down assay was performed to identify that (Arg)9-SH2 superbinder could capture a wide variety of pY proteins. Immunofluorescence was used to detect the efficiency of (Arg)9-SH2 superbinder entering cells. The proliferation ability was assessed by MTT and colony formation assay. In addition, wound healing and transwell assay were performed to evaluate migration of B16F10, A375 and A375/DDP cells. Moreover, apoptosis caused by (Arg)9-SH2 superbinder was analyzed by flow cytometry-based Annexin V/PI. Furthermore, western blot revealed that (Arg)9-SH2 superbinder influenced some pY-related signaling pathways. Finally, B16F10 xenograft model was established to confirm whether (Arg)9-SH2 superbinder could restrain the growth of tumor.ResultsOur data showed that (Arg)9-SH2 superbinder had the ability to enter melanoma cells effectively and displayed strong affinities for various pY proteins. Furthermore, (Arg)9-SH2 superbinder could repress proliferation, migration and induce apoptosis of melanoma cells by regulating PI3K/AKT, MAPK/ERK and JAK/STAT signaling pathways. Importantly, (Arg)9-SH2 superbinder could significantly inhibit the growth of tumor in mice.Conclusions(Arg)9-SH2 superbinder exhibited high affinities for pY proteins, which showed effective anticancer ability by replacing SH2-containing proteins and blocking diverse pY-based pathways. The remarkable ability of (Arg)9-SH2 superbinder to inhibit cancer cell proliferation and tumor growth might open the door to explore the SH2 superbinder as a therapeutic agent for cancer treatment.Electronic supplementary materialThe online version of this article (10.1186/s13046-018-0812-5) contains supplementary material, which is available to authorized users.

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Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis

Cited by 15 publications

References 22 publications

Phosphopeptide enrichment for phosphoproteomic analysis - A tutorial and review of novel materials

Phosphopeptide enrichment for phosphoproteomic analysis - A tutorial and review of novel materials

Combination of Three Functionalized Temperature-Sensitive Chromatographic Materials for Serum Protein Analysis

(Arg)9-SH2 superbinder: a novel promising anticancer therapy to melanoma by blocking phosphotyrosine signaling

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