Catalytic properties of the caspases

Caspase stands for cysteine-dependent aspartate specific protease, and is a term coined to define proteases related to interleukin 1beta converting enzyme and CED-3.1 Thus their enzymatic properties are governed by a dominant specificity for substrates containing Asp, and by the use of a Cys side-chain for catalyzing peptide bond cleavage. The use of a Cys side chain as a nucleophile during peptide bond hydrolysis is common to several protease families. However, the primary specificity for Asp turns out to be very rare among protease families throughout biotic kingdoms. Of all known mammalian proteases only the caspase activator granzyme B, a serine protease, has the same primary specificity. In addition to this unusual primary specificity, caspases are remarkable in that certain of their zymogens have intrinsic proteolytic activity. This latter property is essential to trigger the proteolytic pathways that lead to apoptosis. Here we review the known enzymatic properties of the caspases and their zymogens within the broad context of structure:mechanism:activity relationships of proteases in general.     

Tuning the insertion properties of pHLIP

The pH (low) insertion peptide (pHLIP) has exceptional characteristics: at neutral pH it is an unstructured monomer in solution or when bound to lipid bilayer surfaces, and it inserts across a lipid bilayer as a monomeric alpha-helix at acidic pH. The peptide targets acidic tissues in vivo and may be useful in cancer biology for delivery of imaging or therapeutic molecules to acidic tumors. To find ways to vary its useful properties, we have designed and analyzed pHLIP sequence variants. We find that each of the Asp residues in the transmembrane segment is critical for solubility and pH-dependent membrane insertion of the peptide. Changing both of the Asp residues in the transmembrane segment to Glu, inserting an additional Asp into the transmembrane segment, or replacing either of the Asp residues with Ala leads to aggregation and/or loss of pH-dependent membrane insertion of the peptide. However, variants with either of the Asp residues changed to Glu remained soluble in an aqueous environment and inserted into the membrane at acidic pH with a higher pK_app of membrane insertion.     

Further properties of the two-membrane model

The properties of nonlinear equations describing the solute and solvent transport across a simplified Patlak-Goldstein-Hoffman model (two membranes in series without unstirred layers) are investigated both analytically and numerically. The analysis shows that the principal coefficients measured in transport experiments in the presence of active transport are dependent on the experimental conditions. These ‘apparent’ system parameters are extensions of the corresponding parameters determined both in passive systems and in the linear Kedem-Katchalsky theory. Moreover, they are related to the local phenomenological coefficients of the single membranes of the array. Several relationships between measurable quantities and the local system parameters are indicated, allowing the planning of experiments aimed at the measurement of the latter. Data in the literature have been used to check the proposed volume flow equation.     

On the elastic properties of arteries

A new coefficient of elasticity is proposed that relates to the elastic state of the blood vessels. This measure is proposed as a result of the realization, from personal experience as well as from the international literature, of the difficulty in measuring the thickness of the blood vessels in vivo with acceptable precision. The measurement of E being dependent on the measurement of the thickness of the vessels becomes a highly unreliable proposition. Its relation to E (Young modulus) and to the pulse wave velocity (PWV) is established. We give three examples showing how the proposed coefficient can be measured.