Comparison of the small heat shock proteins alphaB-crystallin, MKBP, HSP25, HSP20, and cvHSP in heart and skeletal muscle

Seven members of the small heat shock protein (sHSP) family are exceptional with respect to their constitutive high abundance in muscle tissue. It has been suggested that sHSPs displaying chaperone-like properties may stabilize myofibrillar proteins during stress conditions and prevent them from loss of function. In the present study five sHSPs (alphaB-crystallin, MKBP, HSP25, HSP20, and cvHSP) were investigated with respect to similarities and differences of their expression in heart and skeletal muscle under normal and ischemic conditions. In ischemic heart and skeletal muscle these five sHSPs translocated from cytosol to the Z-/I-area of myofibrils. Myofibrillar binding of all sHSPs was very tight and resisted for the most part extraction with 1 M NaSCN or 1 M urea. MKBP and HSP20 became extracted by 1 M NaSCN to a significant extent indicating that these two sHSPs may bind partially to actin-associated proteins which were completely extracted by this treatment. Ultrastructural localization of alphaB-crystallin showed diffuse distribution of immunogold label throughout the entire I-band in skeletal muscle fibers whereas in cardiomyocytes alphaB-crystallin was preferentially located at the N-line position of the I-band. These observations indicate different myofibrillar binding sites of alphaB-crystallin in cardiomyocytes versus skeletal muscle fibers. Further differences of the properties of sHSPs could be observed regarding fiber type distribution of sHSPs. Thus sHSPs form a complex stress-response system in striated muscle tissue with some common as well as some distinct functions in different muscle types.     

Neuronal diseases: small heat shock proteins calm your nerves

Mutations in HSPB1 and HSPB8, members of the small heat shock protein family, have recently been shown to cause some distal motor neuropathies. Their function in motor neurones is now under scrutiny.     

Sequences of the mouse N-acetylglucosaminyltransferase V (Mgat5) mRNA and an mRNA expressed by an Mgat5-deficient cell line

N-acetylglucosaminyltransferase V, encoded by the Mgat5 gene, plays an important regulatory role in the synthesis of complex N-glycans, including the Lewis antigens. The elevated expression of this enzyme is regulated during development and is highly associated with cellular transformation and malignancy. In addition, mammary tumors produced in Mgat5 gene knockout mice show markedly reduced metastases, indicating that it plays an important role in this process. The expression of this transferase is important for normal T cell proliferation and function. The amino acid and mRNA sequences for this protein are very highly conserved among the reported species. We report here the sequence of the mouse Mgat5 mRNA. This sequence is very similar to the other mammalian sequences with 95%, 91%, and 87.8% overall sequence identity to rat, hamster, and human mRNAs, respectively. In addition, we have identified a 63-bp insertion mutation, introducing a premature stop codon in the coding region of the Mgat5 mRNA expressed in a N-acetylglucosaminyltransferase V-deficient cell line, PhaR(2.1).     

Insights into the beaded filament of the eye lens

Filensin (BFSP1) and CP49 (BFSP2) represent two members of the IF protein superfamily that are thus far exclusively expressed in the eye lens. Mutations in both proteins cause lens cataract and careful consideration of the detail of these cataract phenotypes alerts us to several interesting features concerning the function of filensin (BFSP1) and CP49 (BFSP2) in the lens. With the first filensin (BFSP1) mutation now having been reported to cause a recessive cataract phenotype, there is the suggestion that the mutation could predispose heterozygote carriers to the early onset of age-related nuclear cataract. In the case of CP49 (BFSP2), there are now three unrelated families who have been identified with a common E233 Delta mutation. Very interestingly this is linked to myopia in one family. Despite the apparent phenotypic differences of the filensin (BFSP1) and CP49 (BFSP2) mutations, the data are still consistent with the beaded filament proteins being essential for lens function and specifically contributing to the optical properties of the lens. The fact that none of the mutations thus far reported affect either the conserved LNDR or TYRKLLEGE motifs that flank the central rod domain supports the view that this pair of IF proteins have unusual structural features and a distinctive assembly mechanism. The multiple sequence divergences suggest these proteins have been adapted to the specific functional requirements of lens fibre cells, a function that can be traced from squid to man.     

The Effect of galE Gene Inactivation on Lipopolysaccharide Profile of Helicobacter pylori

The galE gene product, UDP-galactose 4-epimerase, mediates the incorporation of galactose in extracellular polysaccharide materials such as the O-side chain of lipopolysaccharide (LPS). The O-side chain in H. pylori LPS has been shown to cross-react with Lewis x and/or y blood group antigens, suggesting its potential involvement in H. pylori-linked autoimmune disease. To study its role in H. pylori LPS biosynthesis, the galE gene was cloned, sequenced, and a galE-knockout H. pylori strain was constructed. The H. pylori galE gene encoded a protein of 344 amino acids with a molecular weight of 39K. The LPS profile from the galE-knockout H. pylori strain showed a lower molecular weight than that of the parental strain, indicating the involvement of the galE gene in LPS biosynthesis of H. pylori. Received: 15 December 1997 / Accepted: 10 March 1998     

A comparison of fluorographic methods for the detection of 35S-labeled proteins in polyacrylamide gels

Eight different methods of fluorographic enhancement of sensitivity to 35S decay after gel electrophoresis were compared. Using Kodak X-Omat AR X-ray film, we found that some fluors were about equivalent to 2,5-diphenyloxazole/dimethyl sulfoxide embedding, whereas several other fluors were not quite as effective, but still were significantly more sensitive than control autoradiography. The most sensitive procedures can yield a detectable darkening of film with less than 1 dpm/mm2 of 35S after a 1-week exposure.