Form Of Human Serum Albumin Research Articles

Resistance of cancer cells to immune recognition and killing

It is well recognized that, in order for a wound to heal, the fibrin clot must be eliminated by fibrinolytic enzymes. In certain instances, however, fibrin is ineffectively degraded or even not degraded. For example, in pregnancy, the placenta contains a layer of fibrin (Nitabuck’s layer) which presents as ‘self’ to the immune system. Similar situations have been observed in many solid tumors. A hypothesis is presented according to which tumor cells can escape detection and attack by the immune system in most cancer patients. The tumor dons a ‘coat’ of the host’s own protein on its cell surface. The coat is composed of fibrin and of a polymeric form of human serum albumin (HSA) which, by contrast to pure fibrin, is resistant to fibrinolytic degradation. Such a coated tumor appears as ‘self’ to the immune system, and thus is not detected as a tumor by the immune system (i.e. natural killer cells). When tumors are prepared for in vitro assays against drugs, they are routinely treated with proteolytic enzymes (e.g. pepsin, or chymotrypsin, etc.) which dissolve the protein coat, exposing the tumor cell surface to the drug. Thus, the in vivo existence of a coat on the tumor surface may explain why some drugs have little or no effect in vivo, while the same drugs are active in vitro.

Cross-linked protein crystals for vaccine delivery.

The progress toward subunit vaccines has been limited by their poor immunogenicity and limited stability. To enhance the immune response, subunit vaccines universally require improved adjuvants and delivery vehicles. In the present paper, we propose the use of cross-linked protein crystals (CLPCs) as antigens. We compare the immunogenicity of CLPCs of human serum albumin with that of soluble protein and conclude that there are marked differences in the immune response to the different forms of human serum albumin. Relative to the soluble protein, crystalline forms induce and sustain over almost a 6-month study a 6- to 10-fold increase in antibody titer for highly cross-linked crystals and an approximately 30-fold increase for lightly cross-linked crystals. We hypothesize that the depot effect, the particulate structure of CLPCs, and highly repetitive nature of protein crystals may play roles in the enhanced production of circulating antibodies. Several features of CLPCs, such as their remarkable stability, purity, biodegradability, and ease of manufacturing, make them highly attractive for vaccine formulations. This work paves the way for a systematic study of protein crystallinity and cross-linking on enhancement of humoral and T cell responses.

Determination of the secondary structure of isomeric forms of human serum albumin by a particular frequency deconvolution procedure applied to Fourier transform IR analysis.

A new deconvolution procedure was applied to the analysis of Fourier transform ir spectra of human serum albumin secondary structure in the native state and in states denatured by heat and acid treatment. The deconvolution method is based on the use of the Conjugate Gradient Minimization Algorithm, with the addition of suitable constraints directly obtained by the application to the measured spectrum of the second derivative operator. This method computes central band frequency, bandwidth, and amplitude of the different spectral components of conformation-sensitive amide bands. In the specific case, it was applied to analysis of the amide I band, and the quantitative determination of the different secondary structures (alpha-helix, beta-sheet, beta-turns, and random) was attempted for all the samples examined. The precision of the quantitative determination depends on the amounts of these structures present in the protein. The coefficient of variation is < 10% for values of amide I component > 15%. The accuracy was tested by comparing, by means of linear regression, the results obtained for human serum albumin, hemoglobin, alpha-chymotrypsin, and cytochrome c, using our method, with those obtained by x-ray crystallography and CD; the results obtained by other vibrational spectroscopic approaches were also compared. The fit standard error between x-ray and ir secondary structure values estimated by our method is 2.5% for alpha-helix, 7.16% for beta structures, and 5.1% for other structures (turns and random coils). Quantitative results are given for the secondary structures (alpha-helix, turns, and beta-strands) present in the native state (turns and beta-strands up to now unknown in aqueous solution), together with the percentages of these structures and additional ones (random coils and beta-sheets) formed during denaturization.

5589338 Nucleic acids encoding a mutant form of human serum albumin involved in familial dysalbuminemic hyperthyroxinemia : Bhagavan Nadhipuram; Petersen Charles E; Mandel Morton Honolulu, HI, United States Assigned to University of Hawaii

5589338 Nucleic acids encoding a mutant form of human serum albumin involved in familial dysalbuminemic hyperthyroxinemia : Bhagavan Nadhipuram; Petersen Charles E; Mandel Morton Honolulu, HI, United States Assigned to University of Hawaii

5589338 Nucleic acids encoding a mutant form of human serum albumin involved in familial dysalbuminemic hyperthyroxinemia : Bhagavan Nadhipuram; Petersen Charles E; Mandel Morton Honolulu, United States Assigned to University of Hawaii

5589338 Nucleic acids encoding a mutant form of human serum albumin involved in familial dysalbuminemic hyperthyroxinemia : Bhagavan Nadhipuram; Petersen Charles E; Mandel Morton Honolulu, United States Assigned to University of Hawaii

Increased oxidized form of human serum albumin in patients with diabetes mellitus

High-performance liquid chromatographic (HPLC) analysis of human serum albumin (HSA) on Asahipak GS-520H columns at neutral pH (6.87) showed a clear resolution of human mercaptalbumin (HMA) and nonmercaptalbumin (HNA), which are reduced and oxidized forms of HSA, respectively. We studied the conversion of HMA to HNA (mercapt-nonmercapt conversion) as an index of oxidative change of the tissues and organs in 28 normal subjects and in a total of 47 patients with non-insulin dependent diabetes mellitus (NIDDM). Mean (±SD) values of the HMA fraction of HSA, f(HMA), [ HMA (HMA+HNA) ], was significantly lower in NIDDM patients than in normal subjects (0.63±0.067 vs 0.75±0.028, P<0.001). It was lower in poorly controlled NIDDM patients (0.63±0.058, n=20) than in well controlled NIDDM patients (0.67±0.032, n=9) ( P<0.05). Plasma glucose values sampled on occasions including overnight fasting and postprandial ones ( r= −0.441, n=47, P<0.01), but not plasma glucose values sampled on overnight fasting ( r= −0.345, n=29) or postprandial ( r= −0.467, n=18) conditions and HbAlc ( r= −0.211, n=34), negatively correlated with the f(HMA) values, indicating that mercapt-nonmercapt conversion may not be due to cumulative hyperglycemia over a month, but due to short-term alteration in blood glucose level. The presence or absence of diabetic complications including nephropathy, retinopathy and neuropathy did not affect the f(HMA) values. In conclusion, decreased f(HMA) values in the diabetic patients suggested the presence of a rapidly altered oxidative change of albumin due to hyperglycemia.

Partially folded state of the disulfide-reduced form of human serum albumin as an intermediate for reversible denaturation.

The conformation of the fully disulfide-reduced state of human serum albumin was investigated by tryptophan fluorescence spectrum, CD analyses, and size-exclusion chromatography. Both the reduction of the native disulfide-bonded form under nondenaturing conditions and the refolding of the urea-denatured disulfide-reduced form under reduced conditions yielded almost exactly the same disulfide-reduced state with partially folded unique conformation that was clearly distinguished from either the native or fully denatured state. In addition, the interconversion between the urea-denatured reduced form and the partially folded reduced form was reversible with each other; by reoxidation, the partially folded reduced form was converted to the disulfide-bonded form. The conformation of disulfide-reduced serum albumin was highly variable depending on pH and ionic strength conditions. Thus, we concluded that the disulfide-reduced state with partially folded variable conformation is involved in the reversible interconversion between the denatured reduced form and the native disulfide-bonded form of human serum albumin.

Structural characterization of a chain termination mutant of human serum albumin.

Mutant forms of human serum albumin have been detected on the basis of their abnormal electrophoretic mobility which is either faster or slower than that of normal albumin. In the present work we have studied the structure of a slow variant, referred to as albumin Ge/Ct, in order to define the cause of its genetic abnormality. The protein was isolated from the serum of a young healthy woman homozygous for the variant. Analysis of CNBr fragments by isoelectric focusing allowed us to localize the mutation to the COOH-terminal region of the molecule (residues 549-585). This fragment was isolated on a preparative scale and subjected to tryptic digestion. All tryptic peptides were purified by reverse-phase high performance liquid chromatography and characterized. Sequential analysis of three abnormal peptides revealed that albumin Ge/Ct has a shortened chain with the following COOH-terminal sequence: Leu-Val-Ala-Ala-Ser-Lys580-Leu-Pro. The presence of an additional lysine residue accounts for the electrophoretic behavior of the variant. It is likely that the variant may be caused by a single base deletion in the structural gene, a Cyt in mRNA codon 580, and the consequent shift in reading frame.

The role of the transition between neutral and basic forms of human serum albumin in the kinetics of the binding to warfarin

Between pH 6 and 9 in the kinetics of the binding of warfarin to human serum albumin a two-step mechanism operates: a diffusion-controlled step, followed by a much slower step during which the stable warfarin-albumin complex is formed. The association rate constant for the formation of the warfarin-albumin complex depends on the transition between neutral and basic forms of the albumin.

Letter: X-ray data for four crystalline forms of serum albumin.

Abstract Cell dimensions, space groups and crystal densities are reported for a monoclinic form of human serum albumin (a = 126.5, b = 39.2, c = 135.2A, β = 93.3 °, C2, Z = 4), tetragonal human serum albumin (a = 84.0, c = 276A, P41212, Z = 8), a mercury dimer of human serum albumin (a = 155, b = 83, c = 122A, P212121, Z = 4) and a hexagonal form of equine serum albumin (a = 96.6, c = 144.5A, P61, Z = 6).