Disruption of the AMPK-TBC1D1 nexus increases lipogenic gene expression and causes obesity in mice via promoting IGF1 secretion

Tre-2/USP6, BUB2, cdc16 domain family member 1 (the TBC domain is the GTPase activating protein domain) (TBC1D1) is a Rab GTPase activating protein that is phosphorylated on Ser(231) by the AMP-activated protein kinase (AMPK) in response to intracellular energy stress. However, the in vivo role and importance of this phosphorylation event remains unknown. To address this question, we generated a mouse model harboring a TBC1D1(Ser231Ala) knockin (KI) mutation and found that the KI mice developed obesity on a normal chow diet. Mechanistically, TBC1D1 is located on insulin-like growth factor 1 (IGF1) storage vesicles, and the KI mutation increases endocrinal and paracrinal/autocrinal IGF1 secretion in an Rab8a-dependent manner. Hypersecretion of IGF1 causes increased expression of lipogenic genes via activating the protein kinase B (PKB; also known as Akt)–mammalian target of rapamycin (mTOR) pathway in adipose tissues, which contributes to the development of obesity, diabetes, and hepatic steatosis as the KI mice age. Collectively, these findings demonstrate that the AMPK–TBC1D1 signaling nexus interacts with the PKB–mTOR pathway via IGF1 secretion, which consequently controls expression of lipogenic genes in the adipose tissue. These findings also have implications for drug discovery to combat obesity

AS160 deficiency causes whole-body insulin resistance via composite effects in multiple tissues.

AS160 (Akt substrate of 160 kDa) is a Rab GTPase-activating protein implicated in insulin control of GLUT4 (glucose transporter 4) trafficking. In humans, a truncation mutation (R363X) in one allele of AS160 decreased the expression of the protein and caused severe postprandial hyperinsulinaemia during puberty. To complement the limited studies possible in humans, we generated an AS160-knockout mouse. In wild-type mice, AS160 expression is relatively high in adipose tissue and soleus muscle, low in EDL (extensor digitorum longus) muscle and detectable in liver only after enrichment. Despite having lower blood glucose levels under both fasted and random-fed conditions, the AS160-knockout mice exhibited insulin resistance in both muscle and liver in a euglycaemic clamp study. Consistent with this paradoxical phenotype, basal glucose uptake was higher in AS160-knockout primary adipocytes and normal in isolated soleus muscle, but their insulin-stimulated glucose uptake and overall GLUT4 levels were markedly decreased. In contrast, insulin-stimulated glucose uptake and GLUT4 levels were normal in EDL muscle. The liver also contributes to the AS160-knockout phenotype via hepatic insulin resistance, elevated hepatic expression of phosphoenolpyruvate carboxykinase isoforms and pyruvate intolerance, which are indicative of increased gluconeogenesis. Overall, as well as its catalytic function, AS160 influences expression of other proteins, and its loss deregulates basal and insulin-regulated glucose homoeostasis, not only in tissues that normally express AS160, but also by influencing liver function

The Executable Invocation Policy of Web Services Composition With Petri Net

The web service composition execution engine is a critical problem for web service composition. Based on Petri net and the analysis of structural relationships among web services, the invocation sequence of web service composition and its related invocation policies are fully studied in this paper. Also, the executable invocation policies of web service composition are successfully constructed based on Petri net. Finally, an example of a scientific computation service is given to validate the effectiveness of this method

PKB-Mediated Thr649 Phosphorylation of AS160/TBC1D4 Regulates the R-Wave Amplitude in the Heart.

The Rab GTPase activating protein (RabGAP), AS160/TBC1D4, is an important substrate of protein kinase B (PKB), and regulates insulin-stimulated trafficking of glucose transporter 4. Besides, AS160/TBC1D4 has also been shown to regulate trafficking of many other membrane proteins including FA translocase/CD36 in cardiomyocytes. However, it is not clear whether it plays any role in regulating heart functions in vivo. Here, we found that PKB-mediated phosphorylation of Thr649 on AS160/TBC1D4 represented one of the major PAS-binding signals in the heart in response to insulin. Mutation of Thr649 to a non-phosphorylatable alanine increased the R-wave amplitude in the AS160Thr649Ala knockin mice. However, this knockin mutation did not affect the heart functions under both normal and infarct conditions. Interestingly, myocardial infarction induced the expression of a related RabGAP, TBC1D1, in the infarct zone as well as in the border zone. Together, these data show that the Thr649 phosphorylation of AS160/TBC1D4 plays an important role in the heart's electrical conduction system through regulating the R-wave amplitude

Mechanical Properties of TC11 Titanium Alloy and Graphene Nanoplatelets/TC11 Composites Prepared by Selective Laser Melting

Titanium matrix composites (TMCs) with excellent mechanical properties, reinforced by graphene, is deemed the lightweight and high strength structural materials. In this study, TC11 titanium alloy powder and graphene nanosheets (GNPs) were used as raw materials, and the composite powder with good uniformity and fluidity was obtained through non-interventional homogeneous mixing by a planetary mixer. The microstructure and mechanical properties of the GNPs-TC11 composites and TC11 alloy were compared. The results showed that the microstructure of TC11 and the composites was acicular martensite α’ phase under the process parameters of 280 W laser power, 1200 mm/s scanning speed, and 0.1 mm hatch spacing. The GNPs in addition, in the composites, reduced the acicular martensite particle size and expanded the proportion of low-angle grain boundaries. The tensile strength and percentage elongation after the fracture of the TC11 titanium alloy were 1265 MPa and 4.3%, respectively. Because of addition of the GNPs, the strength and percentage elongation after the fracture of the composite increased to 1384 MPa and 8.1%, respectively, at a GNPs mass content of 0.2%. The enhancement of mechanical properties can be attributed to grain refinement, dislocation strengthening, Orowan strengthening, and load transfer strengthening

The expression of AS160/TBC1D4 and GLUT4 in the four chambers of mouse heart.

<p>A, Immunoblotting analyses of AS160/TBC1D4 and GLUT4 in the lysates (40 μg) of the four chambers of mouse heart. GAPDH was used as a loading control. B, Quantitation of AS160/TBC1D4 expression in the four chambers of mouse heart. n = 6. Asterisk indicates <i>p</i> < 0.05 (atrium versus ventricle). ♯ indicates <i>p</i> < 0.05 (right atrium versus left atrium). C, Quantitation of GLUT4 expression in the four chambers of mouse heart. n = 6. Asterisk indicates <i>p</i> < 0.05 (left ventricle versus the other three chambers).</p

Phosphorylation of AS160/TBC1D4 in mouse heart in response to insulin.

<p>PAS-binding signals, phosphorylation of AS160-Thr⁶⁴⁹ and PKB-Ser⁴⁷³, and total AS160 and PKB were detected in the lysates (40 μg) of hearts from the wild-type and AS160^Thr649Ala knockin mice that were intraperitoneally injected with PBS (basal) or insulin (150 mU per kg of body weight) for 20 min. GAPDH was used as a loading control.</p