Quantifying the Matrix Metalloproteinase 2 (MMP2) Spatially in Tissues by Probe via MALDI Imaging Mass Spectrometry

As a matrix metalloproteinase, the abnormal expression of MMP2 is associated with multiple biological processes, including tissue remodeling and cancer progression. Therefore, spatial analysis of MMP2 protein in tissues can be used as an important approach to evaluate the expression distribution of MMP2 in complex tissue environments, which will help the diagnosis and treatment of various diseases, including tissue or organ injuries. Moreover, this analysis will also help the evaluation of prognoses. However, MMP2 is difficult to be spatially determined by MALDI TOF mass spectrometry due to its large molecular weight (over 72 KD) and low content. Therefore, a new method should be developed to help this detection. Here, we have designed a specific MMP2 probe that closely binds to MMP2 protein in tissue. This probe has a Cl on Tyr at the terminal, which can provide two isotope peaks to help the accuracy quantitative of MMP2 protein. Based on this, we used the probe to determine the spatial expression of MMP2 in the tissues based on MALDI TOF mass spectrometry. This approach may help to study the influence of multifunctional proteases on the degree of malignancy in vivo.

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Quantitative analysis of vitamin D using m/MALDI-TOF mass spectrometry based on a parylene matrix chip

Journal of Analytical Science & Technology ◽

10.1186/s40543-021-00313-2 ◽

2022 ◽

Vol 13 (1) ◽

Author(s):

Joo-Yoon Noh ◽

Moon-Ju Kim ◽

Jong-Min Park ◽

Tae Gyeong Yun ◽

Min-Jung Kang ◽

...

Keyword(s):

Mass Spectrometry ◽

Vitamin D ◽

Quantitative Analysis ◽

Homogeneous Distribution ◽

Fluorescence Image ◽

Real Sample Analysis ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof ◽

Amplex Red ◽

The Matrix

AbstractVitamin D deficiency is associated with various disorders and is diagnosed based on the concentration of 25-hydroxy vitamin D3 (25(OH)D3) in serum. The parylene matrix chip was fabricated to reduce the matrix background noise, and the homogenous distribution of the matrix was retained for the quantitative analysis of 25(OH)D3. The Amplex Red assay was performed to confirm that the sample-matrix mixing zone of the parylene matrix chip was formed below the surface of the parylene-N film. The homogeneous distribution of the matrix was verified from the fluorescence image. For effective analysis using a parylene matrix chip, 25(OH)D3 was modified through the nucleophilic addition of betaine aldehyde (BA) to form a hemiacetal salt. Such modified 25(OH)D3 with a positive charge from BA could be effectively analyzed using MALDI-TOF mass spectrometry. Serum 25(OH)D3 was extracted by liquid–liquid extraction (LLE) and quantified using MALDI-TOF mass spectrometry based on the parylene matrix chip. The intensity of the mass peak of 25(OH)D3 was linearly correlated (r2 = 0.992) with the concentration of 25(OH)D3 spiked in serum, and the LOD was 0.0056 pmol/μL. Energy drinks and vitamin D3 tablets were also employed for the real sample analysis. Finally, the results of the chemiluminescence binding assay and MALDI-TOF mass spectrometry were statistically analyzed to determine the applicability of the method using the Bland–Altman test and Passing–Bablok regression.

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Statins for the prevention of vein graft stenosis: a role for inhibition of matrix metalloproteinase-9

Biochemical Society Transactions ◽

10.1042/bst0300120 ◽

2002 ◽

Vol 30 (2) ◽

pp. 120-126 ◽

Cited By ~ 30

Author(s):

K. E. Porter ◽

N. A. Turner

Keyword(s):

Matrix Metalloproteinase ◽

Intimal Hyperplasia ◽

Therapeutic Potential ◽

Matrix Metalloproteinase 2 ◽

Neointima Formation ◽

Graft Stenosis ◽

The Matrix ◽

And Migration ◽

Proliferation And Migration

Saphenous vein (SV) grafts are commonly used to bypass coronary arteries that are diseased due to atherosclerosis. However, the development of intimal hyperplasia in such grafts can lead to patency-threatening stenosis and re-occlusion of the vessel. The proliferation and migration of smooth muscle cells (SMC) play key roles in the development of intimal hyperplasia, and an agent that inhibits both of these processes therefore has therapeutic potential. A prerequisite for SMC proliferation and migration in vivo is degradation of the basement membrane, achieved by secretion of the matrix-degrading gelatinases matrix metalloproteinase-2 (MMP-2) and MMP-9. Statins are cholesterol-lowering drugs that also have direct effects on SMC function. Here we report that neointima formation in organ-cultured human SV segments is inhibited by simvastatin, an effect that is associated with reduced MMP-9 activity. Additionally, our work shows that simvastatin not only inhibits proliferation, but importantly also inhibits invasion (migration through a matrix barrier), of cultured human SV SMC. Thus simvastatin treatment appears to inhibit neointima formation as a result of combined inhibition of SMC proliferation and invasion. The potential intracellular mechanisms by which statins affect SMC proliferation and migration, and thus attenuate intimal hyperplasia, are discussed, with particular emphasis on the role of MMP-9.

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Probing the in vivo biosynthesis of scytonemin, a cyanobacterial ultraviolet radiation sunscreen, through small scale stable isotope incubation studies and MALDI-TOF mass spectrometry

Bioorganic & Medicinal Chemistry ◽

10.1016/j.bmc.2011.06.005 ◽

2011 ◽

Vol 19 (22) ◽

pp. 6620-6627 ◽

Cited By ~ 11

Author(s):

Carla S. Jones ◽

Eduardo Esquenazi ◽

Pieter C. Dorrestein ◽

William H. Gerwick

Keyword(s):

Mass Spectrometry ◽

Stable Isotope ◽

Ultraviolet Radiation ◽

Small Scale ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof

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Protein Phosphorylation: Sequence-Specific Identification of In Vivo Phosphorylation Sites by MALDI-TOF Mass Spectrometry

Techniques in Protein Chemistry IV ◽

10.1016/b978-0-12-058757-5.50028-7 ◽

1993 ◽

pp. 201-210 ◽

Cited By ~ 6

Author(s):

Tai-Tung Yip ◽

T. William Hutchens

Keyword(s):

Mass Spectrometry ◽

Protein Phosphorylation ◽

Phosphorylation Sites ◽

Specific Identification ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof

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TGF-β Plays a Key Role in Morphogenesis of the Pancreatic Islets of Langerhans by Controlling the Activity of the Matrix Metalloproteinase MMP-2

The Journal of Cell Biology ◽

10.1083/jcb.143.3.827 ◽

1998 ◽

Vol 143 (3) ◽

pp. 827-836 ◽

Cited By ~ 126

Author(s):

Francisco Miralles ◽

Tadej Battelino ◽

Paul Czernichow ◽

Raphael Scharfmann

Keyword(s):

Extracellular Matrix ◽

Matrix Metalloproteinase ◽

Islets Of Langerhans ◽

Neutralizing Antibody ◽

Matrix Metalloproteinase 2 ◽

Pancreatic Islets Of Langerhans ◽

The Matrix ◽

Vitro Model

Islets of Langerhans are microorgans scattered throughout the pancreas, and are responsible for synthesizing and secreting pancreatic hormones. While progress has recently been made concerning cell differentiation of the islets of Langerhans, the mechanism controlling islet morphogenesis is not known. It is thought that these islets are formed by mature cell association, first differentiating in the primitive pancreatic epithelium, then migrating in the extracellular matrix, and finally associating into islets of Langerhans. This mechanism suggests that the extracellular matrix has to be degraded for proper islet morphogenesis. We demonstrated in the present study that during rat pancreatic development, matrix metalloproteinase 2 (MMP-2) is activated in vivo between E17 and E19 when islet morphogenesis occurs. We next demonstrated that when E12.5 pancreatic epithelia develop in vitro, MMP-2 is activated in an in vitro model that recapitulates endocrine pancreas development (Miralles, F., P. Czernichow, and R. Scharfmann. 1998. Development. 125: 1017–1024). On the other hand, islet morphogenesis was impaired when MMP-2 activity was inhibited. We next demonstrated that exogenous TGF-β1 positively controls both islet morphogenesis and MMP-2 activity. Finally, we demonstrated that both islet morphogenesis and MMP-2 activation were abolished in the presence of a pan-specific TGF-β neutralizing antibody. Taken together, these observations demonstrate that in vitro, TGF-β is a key activator of pancreatic MMP-2, and that MMP-2 activity is necessary for islet morphogenesis.

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