AMP-activated protein kinase (AMPK) is

No abstract

To study the structural and functional properties of the human posterior cruciate ligament complex, we measured the cross-sectional shape and area of the anterior cruciate, posterior cruciate, and meniscofemoral ligaments in eight cadaveric knees. The posterior cruciate ligament increased in cross-sectional area from tibia to femur, and the anterior cruciate ligament area decreased from tibia to femur. The meniscofemoral ligaments did not change shape in their course from the lateral meniscus to their femoral insertions. The posterior cruciate ligament cross-sectional area was approximately 50% and 20% greater than that of the anterior cruciate ligament at the femur and tibia, respectively. The meniscofemoral ligaments averaged approximately 22% of the entire cross-sectional area of the posterior cruciate ligament. The insertion sites of the anterior and posterior cruciate ligaments were evaluated. The insertion sites of the anterior and posterior cruciate ligaments were 300% to 500% larger than the cross-section of their respective midsubstances. We determined, through transmission electron microscopy, fibril size within the anterior and posterior cruciate ligament complex from the femur to the tibia. The posterior cruciate ligament becomes increasingly larger from the tibial to the femoral insertions, and the anterior cruciate ligament becomes smaller toward the femoral insertion. We evaluated the biomechanical properties of the femur-posterior cruciate ligament-tibia complex using 14 additional human cadaveric knees. The posterior cruciate ligament was divided into two functional components: the anterolateral, which is taut in knee flexion, and the posteromedial, which is taut in knee extension. The anterolateral component had a significantly greater linear stiffness and ultimate load than both the posteromedial component and meniscofemoral ligaments. The anterolateral component and the meniscofemoral ligaments displayed similar elastic moduli, which were both significantly greater than that of the posteromedial component.

CD22 is a negative regulator of B cell signaling, an activity modulated by its interaction with glycan ligands containing alpha2-6-linked sialic acids. B cells deficient in the enzyme (ST6Gal I) that forms the CD22 ligand show suppressed BCR signaling. Here we report that mice deficient in both CD22 and its ligand (Cd22-/- St6gal1-/- mice) showed restored B cell receptor (BCR) signaling, suggesting that the suppressed signaling of St6gal1-/- cells is mediated through CD22. Coincident with suppressed BCR signaling, B cells lacking ST6Gal I showed a net redistribution of the BCR to clathrin-rich microdomains containing most of the CD22, resulting in a twofold increase in the localization of CD22 together with the BCR. These studies suggest an important function for the CD22-ligand interaction in regulating BCR signaling and microdomain localization.

We recently developed a method for genetically incorporating unnatural amino acids site-specifically into proteins expressed in Escherichia coli in response to the amber nonsense codon. Here we describe the selection of an orthogonal tRNA-TyrRS pair that selectively and efficiently incorporates m-acetyl-l-phenylalanine into proteins in E. coli. We demonstrate that proteins containing m-acetyl-l-phenylalanine or p-acetyl-l-phenylalanine can be selectively labeled with hydrazide derivatives not only in vitro but also in living cells. The labeling reactions are selective and in general proceed with yields of >75%. In specific examples, m-acetyl-l-phenylalanine was substituted for Lys7 of the cytoplasmic protein Z domain, and for Arg200 of the outer membrane protein LamB, and the mutant proteins were selectively labeled with a series of fluorescent dyes. The genetic incorporation of a nonproteinogenic "ketone handle" into proteins provides a powerful tool for the introduction of biophysical probes for the structural and functional analysis of proteins in vitro or in vivo.