Phosphatidylcholine structure determines cholesterol solubility and lipid polymorphism.

In the present work, we demonstrate that phosphatidylcholine with (16:1)9 acyl chains undergoes polymorphic rearrangements in mixtures with 0.6-0.8 mol fraction cholesterol.

The tryptophan-rich region of HIV gp41 and the promotion of cholesterol-rich domains.

The peptide N-acetyl-KWASLWNWFNITNWLWYIK-amide has a sequence that corresponds to the juxtamembrane region of the HIV-1 gp41 fusion protein. We have studied how cholesterol modulates the interaction of this peptide with membranes containing cholesterol using differential scanning calorimetry, circular dichroism, fluorescence spectroscopy, and nuclear magnetic resonance. We find that this peptide is less able to sequester cholesterol into domains than is N-acetyl-LWYIK-amide.

Induction of raft-like domains by a myristoylated NAP-22 peptide and its Tyr mutant.

The N-terminally myristoylated, 19-amino acid peptide, corresponding to the amino terminus of the neuronal protein NAP-22 (NAP-22 peptide) is a naturally occurring peptide that had been shown by fluorescence to cause the sequestering of a Bodipy-labeled PtdIns(4,5)P2 in a cholesterol-dependent manner. The present work, using differential scanning calorimetry (DSC), extends the observation that formation of a PtdIns(4,5)P2-rich domain is cholesterol dependent and shows that it also leads to the formation of a cholesterol-depleted domain. The PtdIns(4,5)P2 used in the present work is extracted from natural sources and does not contain any label and has the native acyl chain composition.

Caveolin scaffolding region and cholesterol-rich domains in membranes.

A protein that constitutes a good marker for a type of cholesterol-rich domain in biological membranes is caveolin. A segment of this protein has a sequence that corresponds to a cholesterol recognition/interaction amino acid consensus (CRAC) motif; this motif has been suggested to cause the incorporation of proteins into cholesterol-rich domains. We have studied the interaction of two peptides containing the CRAC motif of caveolin-1 by differential scanning calorimetry, fluorescence, circular dichroism and magic angle spinning NMR.

Two homologous apolipoprotein AI mimetic peptides. Relationship between membrane interactions and biological activity.

Two related 18-amino acid, class A, amphipathic helical peptides termed 3F-2 and 3F14 were chosen for this study. Although they have identical amino acid compositions and many similar biophysical properties, 3F-2 is more potent than 3F14 as an apolipoprotein AI mimetic peptide. The two peptides exhibit similar gross conformational properties, forming structures of high helical content on a membrane surface.

An apolipoprotein AI mimetic peptide: membrane interactions and the role of cholesterol.

The 18-amino acid amphipathic helical peptide Ac-DWFKAFYDKVAEKFKEAF-NH(2) promotes the separation of cholesterol from the phospholipid, resulting in the formation of cholesterol crystallites, even at mole fractions of cholesterol as low as 0.3. The peptide exerts a greater degree of penetration into membranes of pure phosphatidylcholine in the absence of cholesterol than into bilayers of phosphatidylcholine and cholesterol.

Properties of polyunsaturated phosphatidylcholine membranes in the presence and absence of cholesterol.

Mixtures of cholesterol with phosphatidylcholine species containing the polyunsaturated acyl chains arachidonoyl or docosahexaenoyl were studied by (13)C magic angle spinning (MAS) NMR using both cross-polarization and direct polarization, by (31)P NMR and by differential scanning calorimetry. Several unique features of these systems were observed. The separation of cholesterol in crystalline form occurred at much lower molar fractions than with other forms of phosphatidylcholine.

Peptide-induced formation of cholesterol-rich domains.

The peptide N-acetyl-LWYIK-amide causes the reorganization of bilayers of phosphatidylcholine and cholesterol to produce domains enriched in cholesterol. At a cholesterol mol fraction of 0.5, addition of N-acetyl-LWYIK-amide results in the formation of cholesterol crystallites.

Biosynthesis of vitamin B1 in yeast. Derivation of the pyrimidine unit from pyridoxine and histidine. Intermediacy of urocanic acid.

Incorporation studies with 13C-, 15N-, and 2H-labeled substrates, followed by NMR analysis, show that the pyrimidine unit of thiamin (Vitamin B1) originates from a C5N fragment, derived from C-2',2,N,C-6,5,5' of pyridoxol (Vitamin B6) and an N-C-N fragment derived from L-histidine. Urocanic acid serves as an intermediate on the route of the N-C-N fragment of histidine into the thiamin pyrimidine.

Novel properties of cholesterol-dioleoylphosphatidylcholine mixtures.

We have studied the properties of mixtures of cholesterol with dioleoylphosphatidylcholine (DOPC), and with several other phospholipids, including 1-stearoyl-2-oleoylphosphatidylcholine (SOPC) and dioleoleoylphosphatidylserine (DOPS), as a function of cholesterol molar fraction and of temperature. Mixtures of DOPC with a cholesterol molar fraction of 0.4 or greater display polymorphic behavior.

Biosynthesis of Vitamin B(6) in yeast. Incorporation pattern of trioses.

The biosynthetic origin of the C(3) unit, C-6,5,5', of pyridoxamine was investigated in two yeasts, Candida utilis ATCC 9256 and Saccharomyces cerevisiae ATCC 7752. The incorporation patterns within pyridoxamine bishydrochloride derived from variously multiply (13)C- and (2)H-labeled samples of glycerol and glyceraldehyde, established by NMR spectroscopy, indicate that the three-carbon unit C-6,5,5' of pyridoxamine is derived intact from a triose.

Properties of mixtures of cholesterol with phosphatidylcholine or with phosphatidylserine studied by (13)C magic angle spinning nuclear magnetic resonance.

The behavior of cholesterol is different in mixtures with phosphatidylcholine as compared with phosphatidylserine. In (13)C cross polarization/magic angle spinning nuclear magnetic resonance spectra, resonance peaks of the vinylic carbons of cholesterol are a doublet in samples containing 0.3 or 0.

2'-hydroxypyridoxol, a biosynthetic precursor of vitamins B(6) and B(1) in yeast.

In separate experiments cultures of the yeast Saccharomyces cerevisiae ATTC 7752 were grown in the presence of [5',5'-2H2]- or of [2',2',5',5'-2H4]-3-hydroxy-2,4,5-tri(hydroxymethyl)pyridine (i.e., 2'-hydroxypyridoxol).