Purification and characterization of 26S proteasomes from human and mouse spermatozoa.

Abstract:

:We purified by fractionation on 10-40% glycerol gradients, 26S proteasomes from normal human spermatozoa. These proteasomes, which participate in the ATP-dependent degradation of ubiquitinated proteins, share a similar sedimentation coefficient to those purified from other human tissues. Fluorogenic peptide assays reveal they have chymotrypsin, trypsin and peptidyl-glutamyl-like peptide hydrolysing activities; the chymotrypsin activity is ablated by the specific 26S proteasome inhibitor MG132. Confirmation that these large proteases are 26S proteasomes is provided by detection of the 20S proteasome subunits HC2, XAPC7, RN3 and Z and regulatory ATPases MSS1, TBP1, SUG1 and SUG2 by Western analyses with monoclonal antisera. These antigens are found only in the gradient fractions enriched in proteolytic activities. We have also shown that, although mature spermatozoa from mice have considerably reduced amounts of a ubiquitin-conjugating enzyme (E2) and ubiquitin-protein conjugates in comparison with less mature germ cells, they retain relatively high values of 26S proteasome activity. This suggests that proteasomes may have further roles to play in normal sperm physiology.

journal_name

Mol Hum Reprod

authors

Tipler CP,Hutchon SP,Hendil K,Tanaka K,Fishel S,Mayer RJ

doi

10.1093/molehr/3.12.1053

subject

Has Abstract

pub_date

1997-12-01 00:00:00

pages

1053-60

issue

12

eissn

1360-9947

issn

1460-2407

journal_volume

3

pub_type

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