scholarly journals Baculovirus expression and biochemical characterization of the human microsomal triglyceride transfer protein

1999 ◽  
Vol 338 (2) ◽  
pp. 305-310 ◽  
Author(s):  
Penelope J. RITCHIE ◽  
Anne DECOUT ◽  
Joanna AMEY ◽  
Christopher J. MANN ◽  
Jacqueline READ ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Insect Cells ◽  
Expression System ◽  
Microsomal Triglyceride Transfer Protein ◽  
Low Density Lipoproteins ◽  
Attenuated Total Reflection ◽  
Low Density ◽  
Very Low Density Lipoproteins ◽  
Transfer Protein ◽  
Baculovirus Expression

The microsomal triglyceride transfer protein (MTP) complexed to protein disulphide isomerase (PDI) is obligatory for the assembly of chylomicrons and very-low-density lipoproteins. The determination of the atomic structure of the MTP–PDI heterodimer has important implications for the treatment of those forms of hyperlipidaemia associated with the overproduction of very-low-density lipoproteins, which predispose to premature coronary heart disease. To perform structural studies of the human MTP–PDI complex it was necessary to produce milligram quantities of pure protein. We chose the baculovirus expression system for this purpose. Insects cells were co-infected with recombinant viruses encoding FLAG-tagged MTP and His-tagged PDI; the resulting heterodimer was purified by affinity chromatography. From 5 litres of insect cells, 4–6 mg of more than 95% pure recombinant protein was obtained. CD and attenuated total reflection Fourier-transform infrared spectroscopy indicate that the purified protein has around 34% α-helical and 33% β-structure content. The recombinant protein had a comparable triglyceride transfer activity to that of bovine MTP–PDI. The production of polyclonal antibodies raised against the MTP and PDI subunits of the purified protein is described. The present study demonstrates the feasibility of expressing two proteins at high levels in insect cells and describes a transferable methodology for the purification of the resulting protein complex.

scholarly journals Association of Gene Coding for Microsomal Triglyceride Transfer Protein (MTP) and Meat Texture Characteristic in Pig

2016 ◽  
Vol 16 (3) ◽  
pp. 721-729 ◽  
Author(s):  
Katarzyna Ropka-Molik ◽  
Przemysław Podstawski ◽  
Katarzyna Piórkowska ◽  
Mirosław Tyra
Keyword(s):  
Neutral Lipids ◽  
Membrane Vesicle ◽  
Microsomal Triglyceride Transfer Protein ◽  
Low Density Lipoproteins ◽  
Low Density ◽  
Lipid Transfer ◽  
Texture Characteristic ◽  
Very Low Density Lipoproteins ◽  
Transfer Protein ◽  
Longissimus Lumborum

AbstractThe microsomal triglyceride transfer protein via participation in transport of neutral lipids between membrane vesicle is essential for assembly of chylomicrons, low-density lipoproteins (LDL), and very low-density lipoproteins (VLDL). In human and pigs, it has been confirmed that mutations within MTTP locus affected lipid-transfer activity of this protein. The aim of the present study was to establish potential influences of ENSSSCP00000009789.2:p.Leu840Phe polymorphism on a panel of meat texture parameters measured in two muscles: m. longissimus lumborum and m. semimembranosus. The research performed on 410 pigs showed that investigated missense polymorphism was associated with meat texture profile parameters – TPA (hardness, cohesiveness, springiness, resilience, chewiness) as well as firmness and toughness estimated in loin muscle. In whole analyzed population, the meat of pigs with CC genotype was characterized by significantly the lowest value of TPA characteristic and this trend was also confirmed in two breeds (Puławska and Large White pigs). In turn, the results obtained for firmness and toughness parameters in longissimus lumborum were not consistent across the different populations studied. Our research, in connection with previous studies, indicated that the MTTP gene may be considered as a candidate gene responsible for pork quality traits and pinpointed a need for further analysis in order to select useful genetic markers associated with meat quality parameters.

scholarly journals A Molecular and Biochemical Study for Cholesteryl Ester Transfer Protein (CETP) Taq1B in Iraqi Patients with Hyperlipidemia

2019 ◽  
Vol 16 (3(Suppl.)) ◽  
pp. 0747
Author(s):  
Alkhafajy Et al.
Keyword(s):  
Cholesteryl Ester ◽  
Total Cholesterol ◽  
Cholesteryl Ester Transfer Protein ◽  
Serum Lipid ◽  
Low Density Lipoproteins ◽  
High Density ◽  
High Density Lipoproteins ◽  
Low Density ◽  
Very Low Density Lipoproteins ◽  
Transfer Protein

Cholesteryl ester transfer protein gene contains some single nucleotide polymorphisms, which have been associated with serum high-density lipoprotein concentration and other lipoproteins. This study is done for determining of cholesteryl ester transfer protein polymorphism and evaluate its effect on serum lipid profile concentrations in some hyperlipidemic patients compared with healthy subjects in Salah Al-din governorate-Iraq. Blood samples were taken from (90) patients suffering from hyperlipidemia, and (70) samples that were apparently healthy controls.  Serum lipid concentrations were measured by enzymatic assays. The polymorphism was genotyped using polymerase chain reaction restriction fragment length polymorphism analysis.  The results showed that there was a significant decrease (P<0.05) in the frequency B2 allele, and B1B2, B2B2 genotype, and a significant increase (P<0.05) in the frequency B1 allele, and B1B1 genotype between patients and controls groups. There was a non-significant decrease in the levels of high density lipoproteins, total cholesterol, low density lipoproteins, and very low density lipoproteins levels, and non-significant increase in levels of triglycerides in individuals with the B1B1 genotype than in the B1B2 and B2B2 genotype. However, high density lipoproteins showed a significant decrease (P<0.001) between individuals with the B1B1 genotype and B2B2 genotype. Also, there was a non-significant difference in the levels of high density lipoproteins, total cholesterol, low density lipoproteins, and very low density lipoproteins levels, in individuals with the B1B2 genotype when compared with that of the B2B2 genotype.

scholarly journals Association of missense MTTP gene polymorphism with carcass characteristics and meat quality traits in pigs

2017 ◽  
Vol 62 (No. 1) ◽  
pp. 9-14
Author(s):  
K. Ropka-Molik ◽  
P. Podstawski ◽  
K. Piórkowska ◽  
M. Tyra
Keyword(s):  
Meat Quality ◽  
Microsomal Triglyceride Transfer Protein ◽  
Membrane Vesicles ◽  
Low Density Lipoproteins ◽  
Low Density ◽  
Production Traits ◽  
Protein Activity ◽  
Very Low Density Lipoproteins ◽  
Large White ◽  
Carcass Yield

Microsomal triglyceride transfer protein, coded by MTTP gene, has multiple functions including participation in formation of chylomicrons, low-density lipoproteins, and very low-density lipoproteins. Therefore MTTP protein plays a key role in the transport of fats and cholesterol between membrane vesicles, which can be associated with lipid metabolism. In the present study, ENSSSCT00000010052.2:c.2518C&gt;T (rs335896411) missense polymorphism (Leu&gt;Phe) located in exon 18 of MTTP gene was investigated in order to estimate its potential association with production traits of pigs. The analysis was performed with five breeds (Duroc, Landrace, Large White, Pietrain, Pulawska pigs) and totally 678 pigs, for which the genotypes of c.2518C&gt;T polymorphism were identified by the polymerase chain reaction–restriction fragment length polymorphism method. The present study showed a significant association of c.2518C&gt;T polymorphism with carcass yield. When analyzing the whole population, CC homozygotes showed significantly higher carcass yield than heterozygotes (P ≤ 0.05). Moreover, c.2518C&gt;T single nucleotide polymorphism (SNP) affected pH measured in loin (m. longissimus dorsi) and ham (m. semimembranosus) 45 min after slaughter. For both parameters, the highest pH values were obtained for CC pigs, while the lowest for heterozygotes (P ≤ 0.05). The SNP analyzed was also related with meat colour (yellowness intensity (b*)). Previous research confirmed that ENSSSCP00000009789.2:p.Leu840Phe polymorphism, via affecting MTTP protein activity, influences metabolism of fatty acids. Additionally, results obtained in the present study suggest that the analyzed missense mutation in porcine MTTP gene can be one of the potential genetic factors associated with meat quality (pork pH and colour) and carcass yield.

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