The 43-kD polypeptide of heart gap junctions: immunolocalization, topology, and functional domains

Analysis by SDS-PAGE of gap junction fractions isolated from heart suggests that the junctions are comprised of a protein with an Mr 43,000. Antibodies against the electroeluted protein and a peptide representing the 20 amino terminal residues bind specifically on immunoblots to the 43-kD protein and to the major products arising from proteolysis during isolation. By immunocytochemistry, the protein is found in ventricle and atrium in patterns consistent with the known distribution of gap junctions. Both antibodies bind exclusively to gap junctions in fractions from heart examined by EM after gold labeling. Since only domains of the protein exposed at the cytoplasmic surface should be accessible to antibody, we conclude that the 43-kD protein is assembled in gap junctions with the amino terminus of the molecule exposed on the cytoplasmic side of the bilayer, that is, on the same side as the carboxy terminus as determined previously. By combining proteolysis experiments with data from immunoblotting, we can identify a third cytoplasmic region, a loop of some 4 kD between membrane protected domains. This loop carries an antibody binding site. The protein, if transmembrane, is therefore likely to cross the membrane four times. We have used the same antisera to ascertain if the 43-kD protein is involved in cell-cell communication. The antiserum against the amino terminus blocked dye coupling in 90% of cell pairs tested; the antiserum recognizing epitopes in the cytoplasmic loop and cytoplasmic tail blocked coupling in 75% of cell pairs tested. Preimmune serum and control antibodies (one against MIP and another binding to a cardiac G protein) had no or little effect on dye transfer. Our experimental evidence thus indicates that, in spite of the differences in amino acid sequence, the gap junction proteins in heart and liver share a general organizational plan and that there may be several domains (including the amino terminus) of the molecule that are involved in the control of junctional permeability

AMGEN DRUM HANDLER: LIFTING & TILTING MECHANISM

Team Pick It Up & Put It Down was challenged to design the lifting and tilting portion of a device that will transport chemical-filled plastic barrels from a pallet located on the dirty” side of Amgen’s facilities airlock to another pallet located across a demarcation on the clean” side of the same airlock. This device must also allow for the required 10-minute cleaning process to occur. The device must be reliable, safe, and work in a timely manner to eliminate the current labor-intensive process of manually manipulating these barrels, which can weigh up to 250 pounds. Operation of this device must comply with the particulate standards for an ISO-8 class clean room environment. To meet the design requirements, the team completed thorough market research and review of the 2016-2017 design, assessing the best approach to solving Amgen’s problem. The team used market research and input from Amgen to identify design specifications necessary to develop an effective, quality product. Using brainstorming activities in tandem with patent and literature searches, 120 unique concepts were generated and judged based on their feasibility and effectiveness in solving the given challenge. After careful consideration with the assistance of engineering and concept analysis, it was determined that use of electronics, by means of an electric linear actuator is the most effective concept in eliminating human labor input and abiding by the cleanliness standards. The team worked closely with the University of Rhode Island’s Mechanical Engineering Capstone Team 8, who was challenged to develop the overall device design. Detailed engineering analysis along with side by side development, integrated the lifting and tilting mechanism into a viable concept that functions with the overall design. The team spent the duration of the spring semester building, testing, and redesigning the prototype. At this point in time, the team has effectively developed a product that reduces the physical human manipulation of cleaning the barrels. The clamping and bottom prong system successfully secures barrels during the tilting process when the barrel is correctly placed in contact with the prongs and upper clamp. The team accomplished building an integrated device that improves the safety of day-to-day drum handling and the efficiency of the physical drum positioning for cleaning

The Ursinus Weekly, May 4, 1953

MSGA discusses plagiarism case; Decision rejected • Primary Wed. for M.S.G.A. class officers • Y presidents name cabinet • Sororities elect officers for coming year, 1953-54 • Nominees announced for Curtain Club elections, May 6 • Chem society plans banquet; Officers to be announced • New head waiters chosen • Ursinus-Albright exchange program hears students • Armstrong, Walker are chosen Ruby editors • Pageant, play highlight May Day weekend plans • New MSGA members to be installed, Monday • Retreat combines fun and worship • Doctors speak to pre-meders • Alumni address economics, political science classes • Editorials: Art at last! • Has book-burning begun? • Medical schools report on grads • An open letter praises life in a co-ed college • Belles defeat Rosemont, 4-1 • Experience aids Jayvee tennis • Jayvee court team wins, 4-1 • Trackmen drop trio meet; Lehigh routs Bears, 91-34 • Moravian tops tennis team • Softball team wins over Albright, 35-6 • Ball team wins, 18-4, 4-3; Loses 2-1 to Moravian • Women\u27s Club to sponsor tour of historic county homes • Meistersingers close seasonhttps://digitalcommons.ursinus.edu/weekly/1519/thumbnail.jp