Human lamin B contains a farnesylated cysteine residue
- Univ. of Washington, Seattle (USA)
We recently showed that HeLa cell lamin B is modified by a mevalonic acid derivative. Here we identified the modified amino acid, determined its mode of linkage to the mevalonic acid derivative, and established the derivative's structure. A cysteine residue is modified because experiments with lamin B that had been biosynthetically labeled with ({sup 3}H)mevalonic acid or ({sup 35}S)cysteine and then extensively digested with proteases yielded {sup 3}H- or {sup 35}S-labeled products that co-chromatographed in five successive systems. A thioether linkage rather than a thioester linkage is involved because the mevalonic acid derivative could be released from the {sup 3}H-labeled products in a pentane-extractable form by treatment with Raney nickel but not with methanolic KOH. The derivative is a farnesyl moiety because the Raney nickel-released material was identified as 2,6,10-trimethyl-2,6,10-dodecatriene by a combination of gas chromatography and mass spectrometry. The thioether-modified cysteine residue appears to be located near the carboxyl end of lamin B because treatment of {sup 3}H-labeled lamin B with cyanogen bromide yielded a single labeled polypeptide that mapped toward this end of the cDNA-inferred sequence of human lamin B.
- OSTI ID:
- 5074945
- Journal Information:
- Journal of Biological Chemistry; (USA), Vol. 264:34; ISSN 0021-9258
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PEPTIDE HYDROLASES
ENZYME ACTIVITY
POLYPEPTIDES
MOLECULAR STRUCTURE
AMINO ACID SEQUENCE
CELL NUCLEI
CYSTEINE
GAS CHROMATOGRAPHY
HELA CELLS
MASS SPECTROSCOPY
MEVALONIC ACID
SULFUR 35
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
HYDROGEN COMPOUNDS
HYDROLASES
HYDROXY ACIDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDES
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SPECTROSCOPY
SULFUR ISOTOPES
THIOLS
550201* - Biochemistry- Tracer Techniques