Dissection of De Novo Membrane Insertion Activities of Internal Transmembrane Segments of ATP-Binding-Cassette Transporters: Toward Understanding Topological Rules for Membrane Assembly of Polytopic Membrane Proteins

The membrane assembly of polytopic membrane proteins is a complicated process. Using Chinese hamster P-glycoprotein (Pgp) as a model protein, we investigated this process previously and found that Pgp expresses more than one topology. One of the variations occurs at the transmembrane (TM) domain including TM3 and TM4: TM4 inserts into membranes in an Nin-Cout rather than the predicted Nout-Cin orientation, and TM3 is in cytoplasm rather than the predicted Nin-Cout orientation in the membrane. It is possible that TM4 has a strong activity to initiate the Nin-Cout membrane insertion, leaving TM3 out of the membrane. Here, we tested this hypothesis by expressing TM3 and TM4 in isolated conditions. Our results show that TM3 of Pgp does not have de novo Nin-Cout membrane insertion activity whereas TM4 initiates the Nin-Cout membrane insertion regardless of the presence of TM3. In contrast, TM3 and TM4 of another polytopic membrane protein, cystic fibrosis transmembrane conductance regulator (CFTR), have a similar level of de novo Nin-Cout membrane insertion activity and TM4 of CFTR functions only as a stop-transfer sequence in the presence of TM3. Based on these findings, we propose that 1) the membrane insertion of TM3 and TM4 of Pgp does not follow the sequential model, which predicts that TM3 initiates Nin-Cout membrane insertion whereas TM4 stops the insertion event; and 2) “leaving one TM segment out of the membrane” may be an important folding mechanism for polytopic membrane proteins, and it is regulated by the Nin-Cout membrane insertion activities of the TM segments.

On a common circle: Natural scenes and Gestalt rules

To understand how the human visual system analyzes images, it is essential to know the structure of the visual environment. In particular, natural images display consistent statistical properties that distinguish them from random luminance distributions. We have studied the geometric regularities of oriented elements (edges or line segments) present in an ensemble of visual scenes, asking how much information the presence of a segment in a particular location of the visual scene carries about the presence of a second segment at different relative positions and orientations. We observed strong long-range correlations in the distribution of oriented segments that extend over the whole visual field. We further show that a very simple geometric rule, cocircularity, predicts the arrangement of segments in natural scenes, and that different geometrical arrangements show relevant differences in their scaling properties. Our results show similarities to geometric features of previous physiological and psychophysical studies. We discuss the implications of these findings for theories of early vision.

Toward rules for 1:1 polyamide:DNA recognition

Polyamides composed of four amino acids, imidazole (Im), pyrrole (Py), hydroxypyrrole (Hp), and β-alanine (β), are synthetic ligands that form highly stable complexes in the minor groove of DNA. Although specific pairing rules within the 2:1 motif can be used to distinguish the four Watson⋅Crick base pairs, a comparable recognition code for 1:1 polyamide:DNA complexes had not been described. To set a quantitative baseline for the field, the sequence specificities of Im, Py, Hp, and β for the four Watson⋅Crick base pairs were determined for two polyamides, Im-β-ImPy-β-Im-β-ImPy-β-Dp (1, for Im, Py, and β) and Im-β-ImHp-β-Im-β-ImPy-β-Dp (2, for Hp), in a 1:1 complex within the DNA sequence context 5′-AAAGAGAAGAG-3′. Im residues do not distinguish G,C from A,T but bind all four base pairs with high affinity. Py and β residues exhibit ≥10-fold preference for A,T over G,C base pairs. The Hp residue displays a unique preference for a single A⋅T base pair with an energetic penalty.

An A-DNA triplet code: thermodynamic rules for predicting A- and B-DNA.

The ability to predict macromolecular conformations from sequence and thermodynamic principles has long been coveted but generally has not been achieved. We show that differences in the hydration of DNA surfaces can be used to distinguish between sequences that form A- and B-DNA. From this, a "triplet code" of A-DNA propensities was derived as energetic rules for predicting A-DNA formation. This code correctly predicted > 90% of A- and B-DNA sequences in crystals and correlates with A-DNA formation in solution. Thus, with our previous studies on Z-DNA, we now have a single method to predict the relative stability of sequences in the three standard DNA duplex conformations.

Expansible Computable Aggregation Rules

The aggregation operator have been considered from a computable point of view. The important condition that the computation is friendly when portions of data are inserted o deleted to the list of values to aggregate is considered.

Staying in the Delaware Corporate Governance Lane: Fee Shifting Bylaws and a Legislative Reaffirmation of the Rules of the Road

The Delaware legislature has taken steps towards the adoption of amendments to the Delaware General Corporation Law (DGCL) that would prohibit fee shifting provisions in the articles and bylaws. The language in the legislative proposal, however, addresses fee shifting provisions only in the context of "internal corporate claims." Some have raised concerns that this language would allow for fee shifting provisions that applied to other types of actions, including at least some cases brought under the securities laws. This piece suggests that in fact the Delaware General Corporation Law already prohibits the adoption of bylaws and certificate provisions that apply to causes of action unrelated to internal corporate claims. As a result, there was no reason for the Delaware legislature to expressly bar fee shifting provisions in these types of actions.

Representing Clients in the Marijuana Industry: Navigating State and Federal Rules

As the number of states legalizing medicinal and recreational marijuana increases and marijuana emerges as a growing lawful industry, lawyers find themselves in an awkward position. In most states, lawyers who represent clients in the marijuana industry risk discipline for assisting clients in the commission of a (federal) crime. Even in jurisdictions like Colorado, where the rules of professional conduct have been amended to permit lawyers to assist clients who comply with marijuana state laws, lawyers who are admitted to practice in federal courts risk being disciplined by these tribunals for assisting clients in the commission of a crime pursuant to the courts’ local rules of conduct. This short article explores the thorny issue of navigating state and federal rules of professional conduct while representing clients in the marijuana industry.

Rules and orders respecting the Steward, Cook & Butler of Harvard College, [1686]

These regulations are a revision of the original 1650 rules that laid out the duties of the Steward, Butler, and Cook. In these updated regulations, the Rector and Tutor are now responsible for signing off on the Steward's accounts rather than the President and Fellows.

E. Committee to frame rules, directions, and statutes of the Boylston Professorship of Rhetoric and Oratory, 1804

This subseries consists of a paper notebook containing a handwritten draft of the report presented to the Harvard Corporation on April 30, 1804 by the Committee to frame Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory. The handwritten report provides a numbered list of rules related to the Boylston Professorship and is dated April 16, 1804. The report is followed by a certification signed May 1, 1804 from President Joseph Willard that he was unable to attend the meeting of the Corporation to discuss the professorship.

Draft of the Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory in Harvard College, [ca. 1804]

This original draft was probably written by Eliphalet Pearson (1752-1826) as a member of the committee charged with the task of establishing the rules, directions, and statutes for the Boylston Professorship by the Harvard Corporation and the Board of Overseers. This draft is heavily edited and contains many cross outs through the text.